Expression of Transcription Factors cMyc, Sox2 and klf4 in Human Dental Pulp

Expression of Transcription Factors cMyc, Sox2 and klf4 in Human Dental Pulp | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Expression of Transcription Factors cMyc, Sox2 and klf4 in Human Dental Pulp Heba Alzer, Firas Alsoleihat This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5512553/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: Induced pluripotent stem cells have been proposed as a novel therapeutic option that provides an autologous source of stem cells. However, the methods utilized in generating induced pluripotent stem cells, like viral transduction, ‏and the tumorgenicity of the resulting cells, raise serious concerns in medical society. Objective: Here, we propose that initiating the reprogramming with a type of cell that already has a high expression of some of the transcription factors necessary for the induction would suggestively provide a safer substitute to experiment with for clinical trials. Methods: We investigated the adult human dental pulp cells for the expression of some of these transcription factors, reported previously to induce human somatic cells into pluripotent stem cells. Immunohistochemistry was performed on the wisdom teeth of healthy patients aged between 17 and 19 to study the expression of Sox2, klf4, and c-Myc. Results: The results show a strong expression of Sox2 and c-Myc in the dental pulp among various locations, while klf4 expression was not detected. Conclusion : the human adult dental pulp possesses a subpopulation of stem cells that strongly expresses c-Myc/Sox2 and provides a suitable, feasible source to induce pluripotent stem cells. Stem Cell & Developmental Cell Biology Human embryonic stem cells immunohistochemistry Induced pluripotent stem cells dental pulp transcription factors stem cells. Figures Figure 1 Figure 2 1. Introduction The discovery of induced pluripotent stem cells iPS from adult human dermal fibroblasts formed by retroviral transduction of specific transcription factors represented a landmark in cellular medicine [ 1 ]. Since then, considerable progress has been made in reprogramming technology. Several transcription factors have been examined as potential inducers in numerous transcriptional combinations, including Sox2, c-Myc, and Klf4 [ 1 – 6 ]. Studies have transitioned from dermal fibroblasts to multiple tissue sources such as peripheral blood cells and mesenchymal stem cells in addition [ 4 , 7 ]. Scientists looked for other ways to change cells due to issues with vector transduction. Some methods use small molecules and replacement factors to make cells act differently. These approaches try to prevent genetic and epigenetic changes in the original cells, which is very important for cell transplant therapy [ 2 , 8 – 12 ]. Interestingly, c-Myc, Sox2, and Klf4 are found in human embryonic stem cells. They work together to keep cells in a stem cell state by increasing stemness genes and decreasing differentiation genes [ 7 , 13 ]. Among these developments, our study examines the expression of Sox2, c-Myc, and Klf4 in adult human tooth cells, suggesting that this relatively unexplored niche harbors a capable pluripotent stem cell population with significant potential iPS induction. The authors chose to examine dental pulp native tissue rather than cultured cells, due to concerns that the in vitro culture process may alter the true nature of this subpopulation or prompt it towards differentiation [ 9 , 14 , 15 ]. It should be noted that despite the great promise of dental pulp stem cells, few publications have investigated their characterization by immunohistochemistry, especially with regard to the expression of embryonic stem cell markers. Therefore, the aim of our efforts is to help fill this knowledge gap, to shed light on the expression patterns of key transcription factors within the human dental pulp stem cells, thereby elucidating its potential as a repository of multipotent stem cells. 2. Materials and methods 2.1 Sample collection We collected wisdom teeth from healthy patients aged 17–19 years old. The teeth were extracted for dental reasons, and the study was approved by the Institutional Review Board and Ethics Committee in accordance with The Declaration of Helsinki (decision number 2017/158). Informed consent was obtained to acquire the teeth for analysis, and data privacy was maintained. 2.2 Sample processing After extraction, the teeth were placed in 10% neutral buffered formalin and taken to the laboratory. They were then immersed in a decalcification solution (1:1 volume of 8% formic acid and 8% HCL) and incubated in a water bath at 50°C for 3 hours until they became soft enough to be sectioned. Following this, the teeth were fixed in formalin for 24 hours and then processed in an auto processor, where they underwent fixation in formal saline, dehydration using increasing concentrations of ethanol, clearing with xylene, and finally, infiltration and embedding in paraffin blocks. Using a microtome, serial sections of 3-micrometer thickness were prepared and transferred onto Immunohistochemistry slides. 2.3 Immunohistochemical staining The slides were dried in the dryer for 15 minutes and then incubated overnight at room temperature. After that, they were deparaffinized with xylene for 15 minutes, rehydrated with graded ethanol (ranging from 70–100%), and then washed in distilled water. Heat-mediated antigen retrieval was performed using Tris/EDTA buffer at pH 9 or sodium citrate at pH 6. Table 1 contains the data of the primary antibodies, dilutions, incubation conditions, and antigen retrieval methods used. The remaining steps were carried out exactly as outlined in the EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC Kit [ab80436] by Abcam/UK. Bound antibodies were visualized using a Nikon EcliPSe Ts2 microscope. The location and intensity of the signal were graded as explained in the results in Table 1. Table 1: The primary antibodies utilized for immunohistochemistry. Their clone, source, dilutions, incubation conditions, and antigen retrieval methods. Antibody Clone Source Dilution/ incubation conditions Antigen retrieval method c-Myc Mouse monoclonal [9E10] Abcam UK 1:500 1 hour 25°C Tris/ EDTA pH 9 Sox2 Mouse monoclonal [20G5] Abcam UK 1:200 overnight 4°C Sodium citrate pH 6 KLF4 Rabbit monoclonal [EPR19590] Abcam UK 1:2000 overnight 4°C Tris/ EDTA pH 9 3. Results 3.1 Expression Patterns of Sox2 and c-Myc in Human Dental Pulp Immunohistochemical analysis revealed strong expression of Sox2 and c-Myc in different regions of human dental pulp. High levels of staining of both transcription factors were detected predominantly in the dental pulp tissue, indicating their presence in this niche. Figures 1 and 2 shows the spatial distribution of Sox2 and c-Myc expression within the dental pulp, emphasizing their widespread expression in this tissue. Furthermore, Table 2 provides a comprehensive summary of the strong expression and specific locations where Sox2 and c-Myc were detected in the dental pulp. These observations highlight the relative abundance of these intracellular transcription factors in different cell types of the dental pulp, suggesting a potential role in maintaining cellular pluripotency or regulating cellular functions within this microenvironment. Table 2: The distribution of expression of Sox2, c-Myc, and klf4 in the adult human dental pulp, the intensity of expression is defined in all the locations where the antibodies were detected as follows: +++intense, ++ strong, + moderate, § weak, - negative. Antibody Locations of marker expression Intensity c-Myc Dental pulp core cells Nerve bundles Adipocytes Cell clusters scattered in the dental pulp Sheets in the radicular pulp cell-rich zone Odontoblastic layer - root Odontoblastic layer - crown and cervical part of the root Accessory canals ++ ++ + ++ +++ +++ § +++ Sox2 Dental pulp core cells Cell clusters scattered in the dental pulp Sheets in the radicular pulp cell-rich zone Odontoblastic layer - root Odontoblastic layer - crown and cervical part of the root Subodontoblastic layer Accessory canals ++ +++ +++ +++ +++ + +++ Klf4 Dental pulp - 3.2 Absence of Klf4 Expression in Human Dental Pulp In contrast to the strong expression observed for Sox2 and c-Myc, immunohistochemical analysis did not detect Klf4 expression in human teeth. Despite attempts to detect Klf4 expression using sensitive immunohistochemical techniques, the absence of visible staining suggests either minimal expression levels or complete absence of Klf4. 4. Discussion Sox2, Klf4, and c-Myc are transcription factors expressed in human ESC, they work synergistically to determine pluripotency and self-renewal of human ESC [ 1 – 4 ], and they have been induced in somatic cells to reprogram them into iPS [ 5 , 6 ]. SOX2 regulates self-renewal and odontoblastic differentiation of DPSCs, plays a role in teeth fetal morphogenesis and odontogenesis, and is upregulated in dedifferentiated DPSCs, while Klf4 and c-Myc seem to play a secondary role in early cell fate [ 2 , 7 , 8 ]. Interestingly, the high expression of Klf4 has previously been reported in senescent, terminally differentiated cells [ 9 ], and the expression of c-Myc in most dental pulp stem lines is reported to be low [ 10 ]. To our knowledge, no study has investigated the expression or location of cells expressing these transcription factors in the adult human dental pulp by immunohistochemistry. Earlier reports investigating transcription factors’ expression in in vitro cultures of dental pulp cells reported a low number of DPSCs expressing them [ 8 ]. Moreover, a rapid loss of these transcription factors' expression and the translocation of the signal from the nucleus to the cytoplasm due to cell differentiation are observed [ 8 , 11 ]. However, we showed that c-Myc/Sox2 DPSCs populations exist in the dental pulp in multiple locations [Figures 1 and 2 ]. We suggest that their extinction in in vitro cultures could be attributed to the methods of culture and propagation that caused these cells to differentiate or undergo apoptosis. This is supported by the studies reporting that the enrichment of these DPSCs populations in vitro could be accomplished if DPSCs are cocultured with periodontal ligament stem cells [21], subjected to dedifferentiation [ 17 ] or cultured in neurospheres [ 13 ]. Accordingly, we propose to utilize DPSCs populations expressing c-Myc/Sox2 for iPS induction for the following reasons: Inducing mesenchymal DPSCs into iPS by lentiviral transduction, each carrying one of the four factors Lin28, Nanog, Oct4, and Sox2, produced iPS with a remarkable resemblance to ESC potentials. However, the limitations discussed earlier of viral vector transduction and multiple genetic manipulations still apply [ 10 ]. Moreover, earlier reports showed that forced expression of c-Myc induced differentiation and apoptosis of human ESCs [16, 23] and resulted in a high rate of tumorgenicity among induced cells [ 15 ]. Furthermore, endogenous expression of Sox2 was critical during the induction of somatic cells into iPS [ 4 ]. Many small molecules and replacement factors have been reported to induce Klf4 and oct4 for the completion of the induction of somatic cells into iPS [ 6 , 8 , 9 ]. Accordingly, initiating the induction program with positive c-Myc/Sox2 cells and finalizing the procedure with chemical induction is an option that is worthy of investigating as it could theoretically replace viral induction of genetic material. Further investigations are necessary to optimize the enrichment of these populations in culture before they can be available for future experimentation. Conclusion Dental pulp stem cells are feasible. They would provide a novel source of cells to be induced into iPS. Our results showed that dental pulp contains a population of stem cells that strongly express embryonic transcription factors c-Myc and Sox2 in various locations throughout the dental pulp. We propose utilizing it for iPS induction purposes to minimize the genetic manipulation necessary. Declarations Ethics The Institutional Review Board and Ethics Committee approved the study in accordance with The Declaration of Helsinki, and the decision number is 2017/158. Informed consent was obtained to acquire the teeth for analysis, and data privacy was preserved. Data Availability The data used to publish these findings are available upon request from the authors. Disclaimer Statement The authors have no conflict of interest to announce. Author contribution HA: contributed to conception, design, data acquisition and interpretation, drafted and revised the manuscript. FA: contributed to conception, design, data acquisition and interpretation, and critically revised the manuscript. Funding Statement This study was funded by the Deanship of Scientific Research at the University of Jordan [47/2015/2016]. The funding body played no role in the design of the study and collection, analysis, and interpretation of data nor in writing the manuscript. Acknowledgment The authors would like to thank the Deanship of Scientific Research at the University of Jordan for funding this project, the Maxillofacial surgery department in the University of Jordan Hospital for their cooperation in sample collection, and Mr. Saleh Massad for creating the graphical abstract and preparing the figures. References Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP et al (2005) Core Transcriptional Regulatory Circuitry in Human Embryonic Stem Cells. Cell 122:947–956. https://doi.org/https://doi.org/10.1016/j.cell.2005.08.020 Sumi T, Tsuneyoshi N, Nakatsuji N, Suemori H (2007) Apoptosis and differentiation of human embryonic stem cells induced by sustained activation of c-Myc. Oncogene 26:5564–5576. https://doi.org/10.1038/sj.onc.1210353 González F, Boué S, Belmonte JCI (2011) Methods for making induced pluripotent stem cells: reprogramming à la carte. Nat Rev Genet 12:231–242. https://doi.org/10.1038/nrg2937 Karagiannis P, Takahashi K, Saito M, Yoshida Y, Okita K, Watanabe A et al (2018) Induced Pluripotent Stem Cells and Their Use in Human Models of Disease and Development. Physiol Rev 99:79–114. https://doi.org/10.1152/physrev.00039.2017 Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K et al (2007) Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell 131:861–872. https://doi.org/https://doi.org/10.1016/j.cell.2007.11.019 Velychko S, Adachi K, Kim K-P, Hou Y, MacCarthy CM, Wu G et al (2019) Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs. Cell Stem Cell 25:737–753e4. https://doi.org/https://doi.org/10.1016/j.stem.2019.10.002 Paduano F, Aiello E, Cooper PR, Marrelli B, Makeeva I, Islam M et al (2021) A Dedifferentiation Strategy to Enhance the Osteogenic Potential of Dental Derived Stem Cells. Front Cell Dev Biology ;9 Liu L, Wei X, Ling J, Wu L, Xiao Y (2011) Expression Pattern of Oct-4, Sox2, and c-Myc in the Primary Culture of Human Dental Pulp Derived Cells. J Endod 37:466–472. https://doi.org/https://doi.org/10.1016/j.joen.2010.12.012 Conkright MD, Wani MA, Anderson KP, Lingrel JB (1999) A gene encoding an intestinal-enriched member of the Krüppel-like factor family expressed in intestinal epithelial cells. Nucleic Acids Res 27:1263–1270. https://doi.org/10.1093/nar/27.5.1263 Yan X, Qin H, Qu C, Tuan RS, Shi SHGT (2010) iPS cells reprogrammed from human mesenchymal-like stem/progenitor cells of dental tissue origin. Stem Cells Dev 19:469–480. https://doi.org/10.1089/scd.2009.0314 van Schaijik B, Davis PF, Wickremesekera AC, Tan ST, Itinteang T (2018) Subcellular localisation of the stem cell markers OCT4, SOX2, NANOG, KLF4 and c-MYC in cancer: a review. J Clin Pathol 71 88 LP – 91. https://doi.org/10.1136/jclinpath-2017-204815 Peng Z, Liu L, Zhang W, Wei X (2021) Pluripotency of Dental Pulp Cells and Periodontal Ligament Cells Was Enhanced through Cell-Cell Communication via STAT3/Oct-4/Sox2 Signaling. Stem Cells Int 2021:8898506. https://doi.org/10.1155/2021/8898506 Seo EJ, Jang IH (2020) Increased SOX2 expression in three-dimensional sphere culture of dental pulp stem cells. Korean Acad Oral Biology 45:197–203. https://doi.org/https://doi.org/10.11620/ijob.2020.45.4.197 Cartwright P, McLean C, Sheppard A, Rivett D, Jones K, Dalton S (2005) LIF/STAT3 controls ES cell self-renewal and pluripotency by a Myc-dependent mechanism. Development 132:885–896. https://doi.org/10.1242/dev.01670 Okita K, Ichisaka T, Yamanaka S (2007) Generation of germline-competent induced pluripotent stem cells. Nature 448:313–317. https://doi.org/10.1038/nature05934 Schmidt R, Plath K (2012) The roles of the reprogramming factors Oct4, Sox2 and Klf4 in resetting the somatic cell epigenome during induced pluripotent stem cell generation. Genome Biol 13:251. https://doi.org/10.1186/gb-2012-13-10-251 Li Y, Zhang Q, Yin X, Yang W, Du Y, Hou P et al (2011) Generation of iPSCs from mouse fibroblasts with a single gene, Oct4, and small molecules. Cell Res 21:196–204. https://doi.org/10.1038/cr.2010.142 Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5512553","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":381844588,"identity":"464f0e7f-5559-472c-8cef-5ac3c489e942","order_by":0,"name":"Heba Alzer","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzElEQVRIiWNgGAWjYBACAwYGxgNAWo6xAUjyMDAwE6OFAaTFmHQtiSAdIC2EgTkD84ODPyrupTfPSGB88LaNgZ2fkBbLBjaDAxJninMbZyQwG85tY2CWbCDksANAZNiWANLCJs0L1GJwgKAW9g8HEv8lpDPOSGD/TaQWHoMDBxsSEoBa2JiJ03KYp+Bgw7EEw8aeh82Sc85JEOGX4+0bH/6oSZA3bE8++OFNmU0ywRCDR51hAzgyJZIJ6oADeShtR7yWUTAKRsEoGCkAAMHmPmZyLUyNAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-3841-5418","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Heba","middleName":"","lastName":"Alzer","suffix":""},{"id":381844589,"identity":"5845697e-6426-4951-b801-fb8a4641fa4e","order_by":1,"name":"Firas Alsoleihat","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Firas","middleName":"","lastName":"Alsoleihat","suffix":""}],"badges":[],"createdAt":"2024-11-24 05:55:45","currentVersionCode":1,"declarations":{"humanSubjects":true,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":true,"humanSubjectConsent":true,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-5512553/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5512553/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":69819238,"identity":"7906dca1-aed1-4a1a-bcbb-525a850a8ba0","added_by":"auto","created_at":"2024-11-25 14:01:01","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2376175,"visible":true,"origin":"","legend":"\u003cp\u003eThe distribution of expression of c-Myc in the adult human dental pulp. A] and B] c-Myc was expressed in the dental pulp fibroblasts [black arrows], endothelial cells of venules [white arrows], and the nerve bundles [circles], 20x and 10x, respectively. C] and D] c-Myc was expressed in the dental pulp adipocytes [white arrows] and showed a tendency to cluster in the dental pulp core [circles], both 20x. E] c-Myc heavily positive cells formed sheets in the radicular pulp cell-rich zone [black arrow]. Weak expression is observed in the odontoblastic layer of the cervical area of the root [white arrows] 20x. F] c-Myc was expressed heavily in the radicular odontoblastic layer [white arrows] and the accessory canals [black arrow], 10x. Scale bar 50 um.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5512553/v1/2774758b4072f7ad61603777.png"},{"id":69818173,"identity":"258831ee-49bb-4589-a201-dbe1ddf9765b","added_by":"auto","created_at":"2024-11-25 13:53:01","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":3361321,"visible":true,"origin":"","legend":"\u003cp\u003eThe distribution of expression of Sox2 in the adult human dental pulp. A]\u003c/p\u003e\n\u003cp\u003eSox2 was expressed in the dental pulp fibroblasts [white arrows] and formed clusters in the pulp core [black arrows], 10 x. B] Sox2 heavily positive cells formed sheets in the radicular pulp cell-rich zone [white arrows], 10x. C] and D] many clusters of positive cells were found in the subodontoblastic zone, 20x and 10x, respectively. E] Sox2 was expressed in the radicular odontoblastic layer [black arrows] and the accessory canals [white arrows], 10x. F] Sox2 was expressed heavily in the odontoblastic layer of the crown /cervical area of the root and continued through the accessory canals [black arrow]. Moreover, moderate subodontoblastic zone expression is observed [white arrows], subodontoblastic small clusters are observed [circle], and fibroblasts of the core show an intense expression. 20x. Scale bar 50 um.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5512553/v1/0acda46c9adb3fc7b5b221c4.png"},{"id":69923222,"identity":"cfedb3d5-f985-4fe3-bcfa-2696cd04085f","added_by":"auto","created_at":"2024-11-26 15:52:04","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6611456,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5512553/v1/2ec543bf-3317-4703-8d30-683136d7b654.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eExpression of Transcription Factors cMyc, Sox2 and klf4 in Human Dental Pulp\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe discovery of induced pluripotent stem cells iPS from adult human dermal fibroblasts formed by retroviral transduction of specific transcription factors represented a landmark in cellular medicine [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Since then, considerable progress has been made in reprogramming technology. Several transcription factors have been examined as potential inducers in numerous transcriptional combinations, including Sox2, c-Myc, and Klf4 [\u003cspan additionalcitationids=\"CR2 CR3 CR4 CR5\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Studies have transitioned from dermal fibroblasts to multiple tissue sources such as peripheral blood cells and mesenchymal stem cells in addition [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eScientists looked for other ways to change cells due to issues with vector transduction. Some methods use small molecules and replacement factors to make cells act differently. These approaches try to prevent genetic and epigenetic changes in the original cells, which is very important for cell transplant therapy [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan additionalcitationids=\"CR9 CR10 CR11\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Interestingly, c-Myc, Sox2, and Klf4 are found in human embryonic stem cells. They work together to keep cells in a stem cell state by increasing stemness genes and decreasing differentiation genes [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAmong these developments, our study examines the expression of Sox2, c-Myc, and Klf4 in adult human tooth cells, suggesting that this relatively unexplored niche harbors a capable pluripotent stem cell population with significant potential iPS induction. The authors chose to examine dental pulp native tissue rather than cultured cells, due to concerns that the in vitro culture process may alter the true nature of this subpopulation or prompt it towards differentiation [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIt should be noted that despite the great promise of dental pulp stem cells, few publications have investigated their characterization by immunohistochemistry, especially with regard to the expression of embryonic stem cell markers. Therefore, the aim of our efforts is to help fill this knowledge gap, to shed light on the expression patterns of key transcription factors within the human dental pulp stem cells, thereby elucidating its potential as a repository of multipotent stem cells.\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003e2.1 Sample collection\u003c/h2\u003e\n\u003cp\u003eWe collected wisdom teeth from healthy patients aged 17\u0026ndash;19 years old. The teeth were extracted for dental reasons, and the study was approved by the Institutional Review Board and Ethics Committee in accordance with The Declaration of Helsinki (decision number 2017/158). Informed consent was obtained to acquire the teeth for analysis, and data privacy was maintained.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003e2.2 Sample processing\u003c/h2\u003e\n\u003cp\u003eAfter extraction, the teeth were placed in 10% neutral buffered formalin and taken to the laboratory. They were then immersed in a decalcification solution (1:1 volume of 8% formic acid and 8% HCL) and incubated in a water bath at 50\u0026deg;C for 3 hours until they became soft enough to be sectioned. Following this, the teeth were fixed in formalin for 24 hours and then processed in an auto processor, where they underwent fixation in formal saline, dehydration using increasing concentrations of ethanol, clearing with xylene, and finally, infiltration and embedding in paraffin blocks. Using a microtome, serial sections of 3-micrometer thickness were prepared and transferred onto Immunohistochemistry slides.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003e2.3 Immunohistochemical staining\u003c/h2\u003e\n\u003cp\u003eThe slides were dried in the dryer for 15 minutes and then incubated overnight at room temperature. After that, they were deparaffinized with xylene for 15 minutes, rehydrated with graded ethanol (ranging from 70\u0026ndash;100%), and then washed in distilled water. Heat-mediated antigen retrieval was performed using Tris/EDTA buffer at pH 9 or sodium citrate at pH 6. Table\u0026nbsp;1 contains the data of the primary antibodies, dilutions, incubation conditions, and antigen retrieval methods used. The remaining steps were carried out exactly as outlined in the EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC Kit [ab80436] by Abcam/UK. Bound antibodies were visualized using a Nikon EcliPSe Ts2 microscope. The location and intensity of the signal were graded as explained in the results in Table\u0026nbsp;1.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\n\u003cp\u003e\u003cstrong\u003eTable 1: \u003c/strong\u003eThe primary antibodies utilized for immunohistochemistry. Their clone, source, dilutions, incubation conditions, and antigen retrieval methods.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Taba\" border=\"1\"\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAntibody\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eClone\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSource\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eDilution/ incubation conditions\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAntigen retrieval method\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ec-Myc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMouse monoclonal [9E10]\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAbcam UK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1:500\u003c/p\u003e\n\u003cp\u003e1 hour 25\u0026deg;C\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTris/ EDTA pH 9\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSox2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMouse monoclonal [20G5]\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAbcam UK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1:200\u003c/p\u003e\n\u003cp\u003eovernight 4\u0026deg;C\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSodium citrate pH 6\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKLF4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eRabbit monoclonal [EPR19590]\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAbcam UK\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1:2000\u003c/p\u003e\n\u003cp\u003eovernight 4\u0026deg;C\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTris/ EDTA pH 9\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1 Expression Patterns of Sox2 and c-Myc in Human Dental Pulp\u003c/h2\u003e\n\u003cp\u003eImmunohistochemical analysis revealed strong expression of Sox2 and c-Myc in different regions of human dental pulp. High levels of staining of both transcription factors were detected predominantly in the dental pulp tissue, indicating their presence in this niche. Figures\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e shows the spatial distribution of Sox2 and c-Myc expression within the dental pulp, emphasizing their widespread expression in this tissue. Furthermore, Table\u0026nbsp;2 provides a comprehensive summary of the strong expression and specific locations where Sox2 and c-Myc were detected in the dental pulp. These observations highlight the relative abundance of these intracellular transcription factors in different cell types of the dental pulp, suggesting a potential role in maintaining cellular pluripotency or regulating cellular functions within this microenvironment.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\n\u003cp\u003e\u003cstrong\u003eTable 2:\u0026nbsp; \u003c/strong\u003eThe distribution of expression of Sox2, c-Myc, and klf4 in the adult human dental pulp, the intensity of expression is defined in all the locations where the antibodies were detected as follows: +++intense, ++ strong, + moderate, \u0026sect; weak, - negative.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tabb\" border=\"1\"\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAntibody\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eLocations of marker expression\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eIntensity\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ec-Myc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eDental pulp core cells\u003c/p\u003e\n\u003cp\u003eNerve bundles\u003c/p\u003e\n\u003cp\u003eAdipocytes\u003c/p\u003e\n\u003cp\u003eCell clusters scattered in the dental pulp\u003c/p\u003e\n\u003cp\u003eSheets in the radicular pulp cell-rich zone\u003c/p\u003e\n\u003cp\u003eOdontoblastic layer - root\u003c/p\u003e\n\u003cp\u003eOdontoblastic layer - crown and cervical part of the root\u003c/p\u003e\n\u003cp\u003eAccessory canals\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e++\u003c/p\u003e\n\u003cp\u003e++\u003c/p\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003cp\u003e++\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003cp\u003e\u0026sect;\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSox2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eDental pulp core cells\u003c/p\u003e\n\u003cp\u003eCell clusters scattered in the dental pulp\u003c/p\u003e\n\u003cp\u003eSheets in the radicular pulp cell-rich zone\u003c/p\u003e\n\u003cp\u003eOdontoblastic layer - root\u003c/p\u003e\n\u003cp\u003eOdontoblastic layer - crown and cervical part of the root\u003c/p\u003e\n\u003cp\u003eSubodontoblastic layer\u003c/p\u003e\n\u003cp\u003eAccessory canals\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e++\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003cp\u003e+++\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKlf4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eDental pulp\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2 Absence of Klf4 Expression in Human Dental Pulp\u003c/h2\u003e\n\u003cp\u003eIn contrast to the strong expression observed for Sox2 and c-Myc, immunohistochemical analysis did not detect Klf4 expression in human teeth. Despite attempts to detect Klf4 expression using sensitive immunohistochemical techniques, the absence of visible staining suggests either minimal expression levels or complete absence of Klf4.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eSox2, Klf4, and c-Myc are transcription factors expressed in human ESC, they work synergistically to determine pluripotency and self-renewal of human ESC [\u003cspan additionalcitationids=\"CR2 CR3\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e–\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], and they have been induced in somatic cells to reprogram them into iPS [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. SOX2 regulates self-renewal and odontoblastic differentiation of DPSCs, plays a role in teeth fetal morphogenesis and odontogenesis, and is upregulated in dedifferentiated DPSCs, while Klf4 and c-Myc seem to play a secondary role in early cell fate [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Interestingly, the high expression of Klf4 has previously been reported in senescent, terminally differentiated cells [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], and the expression of c-Myc in most dental pulp stem lines is reported to be low [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo our knowledge, no study has investigated the expression or location of cells expressing these transcription factors in the adult human dental pulp by immunohistochemistry. Earlier reports investigating transcription factors’ expression in \u003cem\u003ein vitro\u003c/em\u003e cultures of dental pulp cells reported a low number of DPSCs expressing them [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Moreover, a rapid loss of these transcription factors' expression and the translocation of the signal from the nucleus to the cytoplasm due to cell differentiation are observed [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. However, we showed that c-Myc/Sox2 DPSCs populations exist in the dental pulp in multiple locations [Figures \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e]. We suggest that their extinction in \u003cem\u003ein vitro\u003c/em\u003e cultures could be attributed to the methods of culture and propagation that caused these cells to differentiate or undergo apoptosis. This is supported by the studies reporting that the enrichment of these DPSCs populations \u003cem\u003ein vitro\u003c/em\u003e could be accomplished if DPSCs are cocultured with periodontal ligament stem cells [21], subjected to dedifferentiation [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] or cultured in neurospheres [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Accordingly, we propose to utilize DPSCs populations expressing c-Myc/Sox2 for iPS induction for the following reasons:\u003c/p\u003e \u003cp\u003eInducing mesenchymal DPSCs into iPS by lentiviral transduction, each carrying one of the four factors Lin28, Nanog, Oct4, and Sox2, produced iPS with a remarkable resemblance to ESC potentials. However, the limitations discussed earlier of viral vector transduction and multiple genetic manipulations still apply [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Moreover, earlier reports showed that forced expression of c-Myc induced differentiation and apoptosis of human ESCs [16, 23] and resulted in a high rate of tumorgenicity among induced cells [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Furthermore, endogenous expression of Sox2 was critical during the induction of somatic cells into iPS [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Many small molecules and replacement factors have been reported to induce Klf4 and oct4 for the completion of the induction of somatic cells into iPS [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Accordingly, initiating the induction program with positive c-Myc/Sox2 cells and finalizing the procedure with chemical induction is an option that is worthy of investigating as it could theoretically replace viral induction of genetic material. Further investigations are necessary to optimize the enrichment of these populations in culture before they can be available for future experimentation.\u003c/p\u003e "},{"header":"Conclusion","content":"\u003cp\u003eDental pulp stem cells are feasible. They would provide a novel source of cells to be induced into iPS. Our results showed that dental pulp contains a population of stem cells that strongly express embryonic transcription factors c-Myc and Sox2 in various locations throughout the dental pulp. We propose utilizing it for iPS induction purposes to minimize the genetic manipulation necessary.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Institutional Review Board and Ethics Committee approved the study in accordance with The Declaration of Helsinki, and the decision number is 2017/158. Informed consent was obtained to acquire the teeth for analysis, and data privacy was preserved.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data used to publish these findings are available upon request from the authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclaimer Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no conflict of interest to announce.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHA: contributed to conception, design, data acquisition and interpretation, drafted and revised the manuscript.\u003c/p\u003e\n\u003cp\u003eFA: contributed to conception, design, data acquisition and interpretation, and critically revised the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Statement\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the Deanship of Scientific Research at the University of Jordan [47/2015/2016]. The funding body played no role in the design of the study and collection, analysis, and interpretation of data nor in writing the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the Deanship of Scientific Research at the University of Jordan for funding this project, the Maxillofacial surgery department in the University of Jordan Hospital for their cooperation in sample collection, and Mr. Saleh Massad for creating the graphical abstract and preparing the figures.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBoyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP et al (2005) Core Transcriptional Regulatory Circuitry in Human Embryonic Stem Cells. 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Cell Res 21:196\u0026ndash;204. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/cr.2010.142\u003c/span\u003e\u003cspan address=\"10.1038/cr.2010.142\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Human embryonic stem cells, immunohistochemistry, Induced pluripotent stem cells, dental pulp, transcription factors, stem cells.","lastPublishedDoi":"10.21203/rs.3.rs-5512553/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5512553/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eInduced pluripotent stem cells have been proposed as a novel therapeutic option that provides an autologous source of stem cells. However, the methods utilized in generating induced pluripotent stem cells, like viral transduction, ‏and the tumorgenicity of the resulting cells, raise serious concerns in medical society.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eObjective: \u003c/strong\u003eHere, we propose that initiating the reprogramming with a type of cell that already has a high expression of some of the transcription factors necessary for the induction would suggestively provide a safer substitute to experiment with for clinical trials.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e We investigated the adult human dental pulp cells for the expression of some of these transcription factors, reported previously to induce human somatic cells into pluripotent stem cells. Immunohistochemistry was performed on the wisdom teeth of healthy patients aged between 17 and 19 to study the expression of Sox2, klf4, and c-Myc.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eThe results show a strong expression of Sox2 and c-Myc in the dental pulp among various locations, while klf4 expression was not detected.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: the human adult dental pulp possesses a subpopulation of stem cells that strongly expresses c-Myc/Sox2 and provides a suitable, feasible source to induce pluripotent stem cells.\u003c/p\u003e","manuscriptTitle":"Expression of Transcription Factors cMyc, Sox2 and klf4 in Human Dental Pulp","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-25 13:52:57","doi":"10.21203/rs.3.rs-5512553/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2ef4a18d-ace2-4244-928d-e6a05246cdd1","owner":[],"postedDate":"November 25th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":40668440,"name":"Stem Cell \u0026 Developmental Cell Biology"}],"tags":[],"updatedAt":"2024-11-25T13:52:57+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-25 13:52:57","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5512553","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5512553","identity":"rs-5512553","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}