In Vitro Expansion of Fetal Liver Hematopoietic Stem Cells

In Vitro Expansion of Fetal Liver Hematopoietic Stem Cells | 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 In Vitro Expansion of Fetal Liver Hematopoietic Stem Cells Rashmi Bhardwaj, Lalit Kumar, Deepika Chhabra, N K Mehra, Atul Sharma, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-156554/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 8 You are reading this latest preprint version Abstract Fetal liver hematopoietic stem cells because of their proliferative potential have been considered appropriate for management of aplastic anemia. Bone marrow recovery was possible in some cases; engraftment potential of these cells however, was unsatisfactory, possibly due to the availability of smaller number of these cells from a single fetus. Present study was undertaken to see if fetal liver hematopoietic stem cells could be expanded in vitro. Mononuclear cells were isolated and hematopoietic stem cells were identified and analyzed by cell surface marker CD34. CD34+ cells were separated by magnetic cell sorting positive selection method using flow cytometry. Hematopoietic stem cells (CD34+) were cultured by using 5 cytokines, stem cell factor, granulocyte macrophages-colony stimulating factor, interlukin-6, Fms-related tyrosine kinase 3 and erythropoietin, in 4 different combinations along with supplements, in serum free culture media for 21 days. Cell viability continued to be greater than 90% throughout 21 days of culture. The cells expanded best in combination of media, supplements and 4 cytokines, namely SCF, Flt-3, IL6 and Epo to yield large number of total (CD34+ & CD34-) cells. Even though the total number of nucleated cells increased in culture significantly, levels of CD34 antigen expression declined steadily over this period. Health Economics & Outcomes Research General Cell Biology & Physiology CD34 + Cell Culture Fetal Liver Hematopoietic stem cells In vitro Figures Figure 1 Figure 2 Figure 3 Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Major revision 16 Feb, 2021 Reviews received at journal 12 Feb, 2021 Reviewers agreed at journal 05 Feb, 2021 Reviewers invited by journal 05 Feb, 2021 Editor assigned by journal 04 Feb, 2021 Editor invited by journal 04 Feb, 2021 Submission checks completed at journal 04 Feb, 2021 First submitted to journal 27 Jan, 2021 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. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-156554","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":10332878,"identity":"73427d72-f4ee-4b79-8430-9dc6d12392e0","order_by":0,"name":"Rashmi Bhardwaj","email":"","orcid":"","institution":"All India Institute of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Rashmi","middleName":"","lastName":"Bhardwaj","suffix":""},{"id":10332879,"identity":"32fd4ecb-be6d-425c-a725-af8106dbfe19","order_by":1,"name":"Lalit Kumar","email":"","orcid":"","institution":"All India Institute of Medical 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Bone marrow recovery was possible in some cases; engraftment potential of these cells however, was unsatisfactory, possibly due to the availability of smaller number of these cells from a single fetus. Present study was undertaken to see if fetal liver hematopoietic stem cells could be expanded in vitro. Mononuclear cells were isolated and hematopoietic stem cells were identified and analyzed by cell surface marker CD34. CD34+ cells were separated by magnetic cell sorting positive selection method using flow cytometry. Hematopoietic stem cells (CD34+) were cultured by using 5 cytokines, stem cell factor, granulocyte macrophages-colony stimulating factor, interlukin-6, Fms-related tyrosine kinase 3 and erythropoietin, in 4 different combinations along with supplements, in serum free culture media for 21 days. Cell viability continued to be greater than 90% throughout 21 days of culture. The cells expanded best in combination of media, supplements and 4 cytokines, namely SCF, Flt-3, IL6 and Epo to yield large number of total (CD34+ \u0026 CD34-) cells. Even though the total number of nucleated cells increased in culture significantly, levels of CD34 antigen expression declined steadily over this period.","manuscriptTitle":"In Vitro Expansion of Fetal Liver Hematopoietic Stem Cells","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-02-08 17:35:58","doi":"10.21203/rs.3.rs-156554/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-02-16T07:35:34+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-02-12T12:32:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"4535c0ec-1eae-4ab1-9606-a339febd4e3b","date":"2021-02-05T15:27:19+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-02-05T15:22:23+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-02-04T13:15:42+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2021-02-04T07:35:30+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2021-02-04T07:32:33+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2021-01-27T09:25:11+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fb527c2a-c00e-45bc-919a-90fecad167d9","owner":[],"postedDate":"February 8th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":2293289,"name":"Health Economics \u0026 Outcomes Research"},{"id":2293290,"name":"General Cell Biology \u0026 Physiology"}],"tags":[],"updatedAt":"2021-04-15T05:44:10+00:00","versionOfRecord":[],"versionCreatedAt":"2021-02-08 17:35:58","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-156554","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-156554","identity":"rs-156554","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}