Effectiveness and Safety of Stem Cell Regeneration for Oocyte Activation in Women with Ovarian Failure: First study using G-CSF mobilization of Peripheral Blood Stem Cells and intraovarian injection of Stem Cell Factor-enriched Platelet Rich Plasma in Clinical Practice

Effectiveness and Safety of Stem Cell Regeneration for Oocyte Activation in Women with Ovarian Failure: First study using G-CSF mobilization of Peripheral Blood Stem Cells and intraovarian injection of Stem Cell Factor-enriched Platelet Rich Plasma in Clinical Practice | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 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Authors Info & Affiliations https://doi.org/10.22541/au.173994868.86444234/v1 453 views 159 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Objective: To evaluate the safety and effectiveness of the Stem Cell Regenera Treatment for oocyte activation in women with ovarian failure in routine clinical practice Design: Retrospective observational study From January 2023 to December 2024 Setting: IVIRMA Alicante Clinics, Spain. Population or Sample: Women diagnosed with ovarian failure including Poor Ovarian Response, Diminished Ovarian Reserve and Premature Ovarian Insufficiency. Methods: The study involved mobilizing Hematopoietic stem cells from bone marrow into peripheral blood using granulocyte colony-stimulating factor, followed by intraovarian injection of Stem Cell Factor-enriched Platelet Rich Plasma. Effectiveness was assessed by correlating leukocytes and CD34+ on day five with oocyte activation criteria and safety assesed by clinical and bilogical parameters. Main Outcome Measures: The primary outcome measures were effectiveness assessed by the rate of oocyte activation, leucocytes and stem cells count, and pregnancy rates. Oocyte activation was defined as an increase in total Antral Follicle Count of three or more follicles after treatment and/or at least a 20% rise in Anti-Müllerian Hormone levels. Safety was assessed based on adverse effects of treatment. Pregnancy rates were evaluated both for spontaneous gestation and following in vitro fertilization treatment. Results: A total of 145 women were included. The overall activation rate was 68.28%, with 7.07% achieving spontaneous gestation and 14.14% achieving pregnancy following IVF. Mobilization of CD34+ cells was successful in all participants, with an average collection of 32.96 CD34+ cells/ µl and no severe adverse effects were observed. Conclusions: The Stem Cell Regenera Treatment is effective and safe for oocyte activation in women with ovarian failure in routine clinical practice Effectiveness and Safety of Stem Cell Regeneration for Oocyte Activation in Women with Ovarian Failure: First study using G-CSF mobilization of Peripheral Blood Stem Cells and intraovarian injection of Stem Cell Factor-enriched Platelet Rich Plasma in Clinical Practice Santamaria A 1 , Ballester A, 2 Muñoz M. 3 1.Hematology Unit. IVIRMA Alicante Clinic. 2. Departamneto de Docencia e Investigación. European University Department, 3. Gynecology Department. IVIRMA Alicante Clinics. Corresponding author: Amparo Santamaria Ortiz. Coordinator Hematology Unit. IVIRMA Alicante Clinic. Email. [email protected] IVIRMA Alicante Clinic. Avenida Denia, Postal Code:03003 Telephone: +34658981769 Abstract Objective: To evaluate the safety and effectiveness of the Stem Cell Regenera Treatment for oocyte activation in women with ovarian failure in routine clinical practice Design: Retrospective observational study From January 2023 to December 2024 Setting: IVIRMA Alicante Clinics, Spain. Population or Sample: Women diagnosed with ovarian failure including Poor Ovarian Response, Diminished Ovarian Reserve and Premature Ovarian Insufficiency. Methods: The study involved mobilizing Hematopoietic stem cells from bone marrow into peripheral blood using granulocyte colony-stimulating factor, followed by intraovarian injection of Stem Cell Factor-enriched Platelet Rich Plasma. Effectiveness was assessed by correlating leukocytes and CD34+ on day five with oocyte activation criteria and safety assesed by clinical and bilogical parameters. Main Outcome Measures: The primary outcome measures were effectiveness assessed by the rate of oocyte activation, leucocytes and stem cells count, and pregnancy rates. Oocyte activation was defined as an increase in total Antral Follicle Count of three or more follicles after treatment and/or at least a 20% rise in Anti-Müllerian Hormone levels. Safety was assessed based on adverse effects of treatment. Pregnancy rates were evaluated both for spontaneous gestation and following in vitro fertilization treatment. Results: A total of 145 women were included. The overall activation rate was 68.28%, with 7.07% achieving spontaneous gestation and 14.14% achieving pregnancy following IVF. Mobilization of CD34+ cells was successful in all participants, with an average collection of 32.96 CD34+ cells/ µl and no severe adverse effects were observed. Conclusions: The Stem Cell Regenera Treatment is effective and safe for oocyte activation in women with ovarian failure in routine clinical practice Funding: No . Keywords: Stem Cell Regenera, oocyte activation, ovarian regeneration, G-CSF, SCFE-PRP, ovarian failure. INTRODUCCIÓN Recent advancements in regenerative medicine have provided new insights into the treatment of ovarian aging and ovarian failure. Women with conditions such as Poor Ovarian Response (POR) and Diminished Ovarian Reserve (DOR) face significant challenges in assisted reproduction. (1-6). These conditions result in a reduced natural conception rate, poor response to stimulation, and limited reproductive potential. Emerging evidence suggests that quiescent follicles in the ovaries can be activated through blood cell-based treatments,(5) including stem cell therapy and Platelet-Rich Plasma (PRP) administration. (7-15) In patients with cancer who undergo chemotherapy and bone marrow transplantation, there have been instances of regained ovarian function and spontaneous pregnancies. This occurrence has been associated with the mobilization of stem cells that reactivate ovarian function (16-19). Research has indicated that various types of stem cells, including mesenchymal, endometrial, and hematopoietic stem cells, have the potential to restore ovarian function (20-22). While most of these studies have been conducted on animal models, some have yielded encouraging results in human subjects. For instance, the ASCOT (Autologous Stem Cell Ovarian Transplantation) study reported notable improvements in ovarian reserve parameters and pregnancy rates following the direct injection of bone marrow stem cells into the ovary. (23) Also, Buigues et al. (24,25) evaluated the noncellular components in different plasma sources enriched with stem cell-secreted factors. They found that plasma from women with POR treated with granulocyte colony-stimulating factor (G-CSF) improved pregnancy rates in mouse models. The regenerative properties of stem cell-enriched plasma were proved in human ovarian tissue, suggesting a paracrine signaling-based alternative for aging ovaries. The experimental pilot study, (26) known as the 4-step-ASCOT technique, evaluated ovarian reserve and reproductive outcomes in POI patients. This study involved mobilizing stem cells from bone marrow into peripheral blood, enriching plasma with stem cell factors, activating platelets, and injecting this enriched plasma directly into the ovaries. The results showed changes in the plasma proteome of POI women, with values like normal responders for a few months. Based on these findings, the StemCell Regenera project was conducted. This treatment combines personalized G-CSF mobilization of Peripheral Blood hematopoietic Stem Cells with intraovarian injection of Stem Cell Factor-enriched (SCFE) PRP. The study aimed to assess the safety and effectiveness of the StemCell Regenera Treatment for oocyte activation and ovarian regeneration in routine clinical practice. MATERIAL Y MÉTODOS From January 2023 to December 2024, we conducted a retrospective observational study of women diagnosed with Poor ovarian response (POR) and Diminished Ovarian Reserve (DOR), POI (premature ovarian insufficiency). As inclusion criteria, we included women with Diminished Ovarian Reserve (DOR), Poor Ovarian Response (POR), and Premature Ovarian Insufficiency (POI). DOR refers to the decrease in ovarian reserve, with diagnostic criteria defined by Poseidon Groups 3 and 4. Group Poseidon 3 includes women under 35 years old with low ovarian reserve, characterized by Anti-Müllerian Hormone (AMH) levels below 1.2 ng/mL and an Antral Follicular Count (AFC) of fewer than 5 follicles, regardless of previous ovarian stimulation response. Group Poseidon 4 includes women aged 35 years or older with low ovarian reserve, also defined by AMH levels below 1.2 ng/mL and AFC of fewer than 5 follicles, regardless of previous ovarian stimulation response. POR is diagnosed according to the Bologna classification by ESHRE, which includes women aged 40 years or older or those with any indication of reduced ovarian reserve (low AMH, high FSH, low AFC) and a poor response in a previous IVF cycle (≤3 oocytes retrieved with standard stimulation). POI includes women who meet the ESHRE criteria for POI, characterized by the presence of menstrual disturbance defined as oligo/amenorrhea for at least 4 months, biochemical confirmation as evidenced by an elevated FSH level fluctuating POI when one of the above criteria is missing. The exclusion criteria were age under 18, genetic or autoimmune ovarian insufficiency, clinical endometriosis, previous abdominal surgeries causing pelvic adhesions, non-visible ovaries inaccessible vaginally, positive serologies for HBsAg, HBcAc, HCV, HIV, syphilis, significant abnormalities in analytical results, and known intolerance or allergies to study product components. Methods This study was conducted from January 2023 to December 2024 and involved women diagnosed with Poor Ovarian Response (POR), Diminished Ovarian Reserve (DOR), and Premature Ovarian Insufficiency (POI). The primary objective was to gather leukocytes and CD34+ cells for oocyte activation and examine their correlation with activation criteria. Effectiveness was measured by linking leukocytes and CD34+ cells on day five to the activation criteria, defined as a positive response when women showed an increase in Antral Follicle Count (AFC) by ultrasound of three or more follicles after treatment or at least a 20% rise in Anti-Müllerian Hormone (AMH) levels. The primary objective was to gather leukocytes and CD34+ cells for oocyte activation and examine their correlation with activation criteria. Effectiveness was measured by linking leukocytes and CD34+ cells on day five to the activation criteria, defined as a positive response when women showed an increase in Antral Follicle Count (AFC) by ultrasound of three or more follicles after treatment or at least a 20% rise in Anti-Müllerian Hormone (AMH) levels. Safety was determined based on adverse effects from G-CSF treatment, categorized as follows: none, mild (mild headache or bone pain without the need for pain medication), moderate (bone pain, headache, or other symptoms related to G-CSF requiring pain treatment or cessation of medication for 24 hours), and severe (bone pain or other related symptoms requiring intravenous pain treatment that led to discontinuation of G-CSF treatment). The secondary objectives included evaluating the analytical parameters of each patient before and after G-CSF treatment. This involved quantifying red and white blood cells and platelets before and after treatment, measuring CD34+ prior to the PRP intraovarian injection, assessing follicular development through antral follicle count, determining the number of activated oocytes, and calculating the pregnancy rate The study procedures were approved by the Institutional Review Board of Hospital Universitario y Politécnico La Fe, Valencia, Spain, and the Ethics Committee from the University of Valencia, Valencia, Spain. Written informed consent was obtained from all participants. These women chose the Stem Cell Regenera Treatment instead of oocyte donation at IVIRMA Alicante Clinics, Spain. All study procedures were approved and conducted according to the Institutional Review Board of Hospital Universitario y Politécnico La Fe, Valencia, Spain and the Institutional Review Board and the Ethics Committee from University of Valencia, Valencia, Spain (2403-ALC-039-AS). Written Informed consent was obtained. The Stem Cell Regenera Treatment comprises a tailored mobilization of stem cells into the bloodstream using granulocyte colony-stimulating factor (G-CSF), followed by the intraovarian injection of SCFE PRP. The tailored mobilization protocol involves adjusting the G-CSF dosage daily based on leukocyte count and clinical symptoms reported by the patient. This customization ensures optimal mobilization of hematopoietic stem cells while minimizing adverse effects and maximizing the therapeutic benefits. Figure 1 illustrates the schedule for the SC Regenera Treatment. The first stage of the treatment was four-day mobilization with G-CSF Injection, starting with an SC dose of 10 mg/kg/d, adjusting daily based on leukocyte control. On the fifth day, stem cell circulating count and white blood cells were performed. Samples were analyzed by flow cytometry to quantify the CD34+ CD45+ population in the flow cytometer (DxFLex). Figure 1 . Schedule for the SC Regenera Treatment. The second stage of treatment involved injecting SCFE-PRP into the ovarian marrow. The treatment consisted of an intraovarian injection of SCFE-PRP; administration to both ovaries in an ultrasound-guided process like that of an oocyte puncture, on the fifth day. The procedure to obtain SCFE-PRP-Endoret was according to a medical device developed by BTI Biotechnology Institute. (BTI®), the PRP was prepared according to a standard protocol (Endoret kit; BTI®). The final sample obtained was quantified to verify the platelet concentration and was injected transvaginally distributed equally between both ovaries. The total amount of SCFE -PRP injected ranged between 3.6 and 8 ml1.) (9) Statistical analysis Statistical analysis was conducted using Chi-square and T-tests were used for median and range comparisons, nonparametric ROC curves and tests for equality of ROC areas across different age groups. Logistic regression was performed as well as Pearson Correlation. A p-value greater than 0.05 was considered statistically significant. RESULTS We recruited 265 women for the study, and 182 met the inclusion criteria. However, follow-up data was lost for 37 women. See Figure 1. Therefore, a total of 145 women were included in the final analysis, with an average age of 39 years (ranging from 26 to 44 years). The mean age was 39,1 years old. and ranged from 26 to 44 years, most participants identified as Caucasian (77.2%), followed by Arab (15.2%), Hispanic (4.8%), and other ethnicities. The initial platelet count was 247,910 x 10 12 /mm3 (range:107,000 - 455,000.) The mean white blood cell count was 6,025 x 10 9 /mm3 (range of 3,010 -10,220. x10 9 /mm3.and the mean hemoglobin was 13 g/dl (range 7.5-18.2). Demographic and characteristics of women are presented in Table 1. Figure 2. Flow Chart depicting the participant selection, inclusion criteria, and outcomes in the StemCell Regenera Study. Table 1. Characteristic and Demographic data depending on Oocyte activation. No Yes P-Value Age (Years-old, Range) 40 (26-44) 40 (31-44) 0.9 Hemoglobin at baseline(gr/dL) Mean, Range 13.1 (7.6-15.8) 13.6 (10,4-18,2) 0.2 Leucocytes at baseline (mm3) Mean, Range 5752 (3290-9900) 6151 (3010-10220) 0.2 Mean Platelets at baseline (mm3), Mean, Range 252522 (107000-455000) 245768 (150000-403000) 0.5 Effectiveness of the Stem Cell Regenera The Stem Cell Regenera Study aimed to evaluate the effectiveness and safety of stem cell mobilization using granulocyte colony-stimulating factor (G-CSF) and intraovarian injection of stem cell factor-enriched platelet-rich plasma (SCFE-PRP) for oocyte activation in women with poor ovarian response (POR) and diminished ovarian reserve (DOR).The age distribution of the patients was as follows: the mean age was 39.1 years, ranging from 26 to 44 years. Most participants identified as Caucasian (77.2%), followed by Arab (15.2%), Hispanic (4.8%), and other ethnicities 1. When comparing the age distribution between activated and non-activated groups, there was no significant difference in age (p=0.9). The overall oocyte activation rate was 68.28% of patients achieved OA (p<0.001) and the gestation rates observed were as follows: 7.07% of activated patients achieved spontaneous gestation (n=7), with a statistical significance of p<0.001. The overall gestation rate was 14.14% following treatment IVF (n=14), also with a statistical significance of p<0.001. None of the women without activation achieved pregnancy, either spontaneously or through IVF. See Table 2. Table 2. Outcomes of Stem cell Regenera in OA and Gestational Rate and Leucocytes and CD34+ on fifth day. * P<0.00 ** P<0.5 *** P<0.05 No 46 31.72 0 0 37666 (20660- 58250) 26,5 (9-73) Yes 99 68.28 7 (7.07) 14 (14.14) 36359 (14470-59670) 33,1 (2-140) G-CSF Mobilization Effectiveness Evaluation Stem Cell Mobilization : Mobilization of CD34+ cells was successful in all participants, with an average collection of 3000 CD34+ cells/ mm3 The median number of CD34+ cells in women who achieved oocyte activation was slightly higher than in those who did not (33.1 vs. 26.5, p=0.05). The median leukocyte count in women with oocyte activation was 36,359 x mm 3 (range: 14,470-59,670). During the four-day treatment, nearly 30% of women needed adjustments to their G-CSF dosage. This depended on their white blood cell count or the presence of moderate symptoms to achieve sufficient stem cell collection with minimal discomfort. Therefore, strict and daily monitoring was conducted. The effectiveness was analyzed by age groups, older and younger than 40 years old, and different diagnosis, but no differences were seen between either groups. The Pearson correlation coefficient was 0.465, showing a moderate positive relationship between white blood cells and CD34+ cells/µl. This suggests that as the value of white blood cells increases, the value of CD34+ cells/µl also tends to increase, with a statistical significance (p < 0.001) suggesting that this correlation is highly unlikely to be due to chance. In a clinical context, this moderate positive correlation implies that there is a consistent and meaningful relationship between the two variables. This can be useful for understanding how these variables behave together and can inform monitoring or treatment decisions. The significant correlation shows that the relationship is reliable and not random, which adds confidence to its clinical relevance. The study aimed to establish a cutoff for CD34+ cells and leukocytes corresponding to a high rate of oocyte activation by analyzing different age groups and cutoff values. We made Women under 38 years of age exhibited a higher rate of activation compared to those aged 38 years and older. The distribution of CD34+ values between these two age groups showed no significant difference, and similarly, the distribution of leukocyte values did not present a significant difference between the groups. A leukocyte count of 33,340 / mm3 was identified as the optimal cutoff point to maximize the difference between the true positive rate (TPR) and the false positive rate (FPR). Additionally, a CD34+ value of 5248.8 / mm3 was determined to be the best cutoff point for maximizing the difference between TPR and FPR. However, the area under the curve value of 0.487 indicates that CD34+ counts have a low capacity to differentiate between activation states in this study, potentially due to the small number of patients or other associated variables, or it may suggest that there is no definitive cutoff predicting activation. Regression analysis suggested that variables such as age, leukocytes, and CD34+ significantly influence activation. Nonetheless, the model’s performance, given the limited patient sample, implies that other unconsidered factors like the patient’s baseline health, ovarian reserve, treatment protocol, and dosage of granulocyte colony-stimulating factor (G-CSF) may also contribute to the outcomes and require further statistical validation. G-CSF Mobilization Safety Evaluation Adverse Effects : Most participants experienced only mild side effects, with no serious adverse effects reported. Specifically, 53.1% of women showed no symptoms, 32.8% experienced mild effects, and 14.1% encountered moderate to severe effects. Following mobilization treatment, all participants showed normalization of their peripheral blood counts. See Table 3. Symptom Observation : Daily observation of symptoms showed that most women experienced either no symptoms or only mild symptoms. The proportion of women experiencing mild symptoms increased from 8.28% on Day +1 to 58.6% on Day +4, while moderate symptoms were less common, and no symptoms decreased over time. See Table 3. Table 3. Safety of the mobilization in terms of side effects associated with the treatment. Mild 12 (8.28%) 59 (40.69%) 72 (49.66%) 85 (58.62%) Moderate 0 12 (8.28%) 20 (13.79%) 17 (11.72%) None 74 (51.03%) 74 (51.03%) 53 (36.55%) 43 (29.66%) Main Findings This retrospective observational study evaluated the safety and effectiveness of the StemCell Regenera treatment for oocyte activation in 145 women diagnosed with ovarian failure, encompassing Poor Ovarian Response (POR), Diminished Ovarian Reserve (DOR), and Premature Ovarian Insufficiency (POI). The main finding was that 68.28% of patients achieved oocyte activation, defined as either an increase in total Antral Follicle Count (AFC) of three or more follicles after treatment or at least a 20% rise in Anti-Müllerian Hormone (AMH) levels. Furthermore, 7.07% of activated patients achieved spontaneous gestation, and 14.14% achieved pregnancy following IVF. Mobilization of CD34+ cells was successful in all participants, with an average collection of 3000 CD34+ cells/mm3, and no severe adverse effects were observed. A trend towards a higher median number of CD34+ cells in women who achieved oocyte activation compared to those who did not reach borderline statistical significance (p=0.05). Strengths and Limitations The strengths of this study include its evaluation of a relatively large cohort of women (n=145) undergoing StemCell Regenera treatment in a real-world clinical setting. The use of a standardized treatment protocol, involving G-CSF mobilization of peripheral blood hematopoietic stem cells and intraovarian injection of SCFE-PRP, allows for consistent assessment of outcomes. Furthermore, the clear definition of oocyte activation based on AFC and AMH levels provides objective measures of treatment effectiveness. The successful mobilization of CD34+ cells in all participants, coupled with the absence of severe adverse effects, underscores the safety of the treatment protocol. The primary limitation of this study is its retrospective observational design, which does not allow for establishing a causal relationship between the StemCell Regenera treatment and the observed improvements in oocyte activation and pregnancy rates. The lack of a control group makes it difficult to discern whether the observed outcomes are directly attributable to the treatment or reflect spontaneous fluctuations in ovarian function. The patient population was heterogeneous in terms of the underlying cause of ovarian failure (POR, DOR, POI), which may introduce variability in treatment response. Data on the number of previous IVF attempts, a factor known to influence pregnancy outcomes, was not available. The absence of detailed information on semen quality parameters, fertilization rates, embryo quality, and live birth rates further limits the interpretability of the results. Moreover, as stated in the exclusion criteria, those with genetic causes of POI were excluded; therefore, this stem cell regeneration may not be an option for those cases. Lastly, the lack of long-term follow-up data precludes assessment of the durability of treatment effects and the potential for adverse outcomes. Interpretation (in light of other evidence) The findings of this study align with previous research suggesting that stem cell-based therapies may improve ovarian function in women with ovarian failure. For example, the ASCOT study reported improvements in ovarian reserve parameters and pregnancy rates following the direct injection of bone marrow stem cells into the ovary (23). The StemCell Regenera treatment shares similarities with the 4-step-ASCOT technique (26), which involves mobilizing stem cells, enriching plasma with stem cell factors, activating platelets, and injecting the enriched plasma directly into the ovaries [14,20-22) While the StemCell Regenera treatment mobilizes peripheral blood stem cells rather than bone marrow stem cells and uses a tailored G-CSF mobilization protocol, the underlying principle of promoting ovarian regeneration through stem cell-mediated mechanisms is still the same. However, the results of this study should be interpreted in the context of the existing literature, which includes conflicting findings about the effectiveness of stem cell-based therapies for ovarian failure. While some studies have reported promising results, others have found no significant benefit or even adverse outcomes [1-15, 23-24). The heterogeneity of patient populations, treatment protocols, outcome measures, and study designs may account for these discrepancies. The finding that a higher median number of CD34+ cells was seen in women with oocyte activation, while reaching borderline statistical significance, it suggests that the mobilization of stem cells may play a role in the treatment’s effectiveness. This is consistent with earlier research showing that CD34+ cells, a subset of hematopoietic stem cells, can promote angiogenesis and tissue regeneration (14) .However, the lack of statistical significance underscores the need for further investigation to elucidate the specific mechanisms through which stem cells may improve ovarian function. In comparison to these studies, Buigues et al. (26) evaluated the noncellular components in different plasma sources enriched with stem cell-secreted factors. They found that plasma from women with POR treated with granulocyte colony-stimulating factor (G-CSF) improved pregnancy rates in mouse models. The regenerative properties of stem cell-enriched plasma were proved in human ovarian tissue, suggesting a paracrine signaling-based alternative for aging ovaries. Compared to oocyte donation, which is a therapeutic option for many women with ovarian failure, the StemCell Regenera treatment offers the potential to use a patient’s own genetic material, which can have significant psychological benefits. However, patients should be counseled about the limited evidence about its effectiveness and safety, as well as the potential for adverse outcomes. Conclusion The StemCell Regenera treatment, combining G-CSF mobilization of peripheral blood stem cells and intraovarian injection of SCFE-PRP, appears to be effective and safe for oocyte activation in women with ovarian failure. This treatment protocol mobilized CD34+ cells in all participants with no severe adverse effects, suggesting that stem cells and SCFE PRP may play a role in the treatment’s effectiveness. The achievement of spontaneous gestations and pregnancies following IVF is an encouraging outcome, but further research is needed to confirm the findings of this study. Acknowledgements Thank you to all the staff in the clinics, and also to the patients Disclosure of Interests: none Contribution to Authorship A.S. contributed to the study’s conception, planning, design, analysis, writing, review, and final approval. AB was involved in the design, recruitment, writing, review, and final approval. M.M. (Muñoz M) participated in the design, analysis, review, and final approval. Each author accepts responsibility for the published paper. Details of Ethics Approval: The study procedures were approved by the Institutional Review Board of Hospital Universitario y Politécnico La Fe, Valencia, Spain, and the Ethics Committee from the University of Valencia, Valencia, Spain. Written informed consent was obtained from all participants. References 1. Na J, Kim GJ. Recent trends in stem cell therapy for premature ovarian insufficiency and its therapeutic potential: a review. J Ovarian Res. 2020;13(1):74. doi: 10.1186/s13048-020-00671-2. 2. Ciccocioppo R, Cantore A, Chaimov D, Orlando G. Regenerative medicine: the red planet for clinicians. Intern Emerg Med. 2019;14(6):911-921. doi: 10.1007/s11739-019-02126-z. 3. Ferraretti AP, La Marca A, Fauser BC, Tarlatzis B, Nargund G, Gianaroli L, and ESHRE working group on Poor Ovarian Response Definition. ESHRE consensus on the definition of ’poor response’ to ovarian stimulation for in vitro fertilization: the Bologna criteria. Hum Reprod. 2011; 26:1616–24. https://doi.org/10.1093/humrep/der092 PMID:21505041 4. Webber L, Davies M, Anderson R, Bartlett J, Braat D, Cartwright B, Cifkova R, de Muinck Keizer-Schrama S, Hogervorst E, Janse F, Liao L, Vlaisavljevic V, Zillikens C, Vermeulen N, and European Society for Human Reproduction and Embryology (ESHRE) Guideline Group on POI. ESHRE Guideline: management of women with premature ovarian insufficiency. Hum Reprod. 2016; 31:926–37. https://doi.org/10.1093/humrep/dew027 PMID:27008889 5. van Kasteren YM, Schoemaker J. Premature ovarian failure: a systematic review on therapeutic interventions to restore ovarian function and achieve pregnancy. Hum Reprod Update. 1999;5(5):483-492. 6. Hansen KR, Craig LB, Zavy MT, Klein NA, Soules MR. Ovarian primordial and non-growing follicle counts according to the Stages of Reproductive Aging Workshop (STRAW) staging system. Menopause. 2012;19(2):164. 7. Huang Q, et al. G-CSF-mobilized peripheral blood mononuclear cells combined with platelet-rich plasma accelerate restoration of ovarian function in cyclophosphamide-induced POI rats. Biol Reprod. 2019;101(1):91-101. 8. Pantos K, Simopoulou M, Pantou A, Rapani A, Tsioulou P, Nitsos N, et al. A case series on natural conceptions resulting in ongoing pregnancies in menopausal and prematurely menopausal women following platelet-rich plasma treatment. Cell Transplant. 2019;28(9-10):1333-1340. 9. Cantero MM. Not all platelet-rich plasma are created equal. Curr Opin Obstet Gynecol. 2024 Jun 1;36(3):118-123. 10. Díaz-García C, Herraiz S, Pamplona L, Subirá J, Soriano MJ, Simon C, et al. Follicular activation in women previously diagnosed with poor ovarian response: a randomized, controlled trial. Fertil Steril. 2022;117(4):747-755. 11. Herraiz S, et al. Treatment potential of bone marrow-derived stem cells in women with diminished ovarian reserves and premature ovarian failure. Curr Opin Obstet Gynecol. 2019;31(3):156-162. 12. Shareghi-oskoue O, Aghebati-Maleki, L. & Yousefi, M. Transplantation of human umbilical cord mesenchymal stem cells to treat premature ovarian failure. Stem Cell Res Ther 12 , 454 (2021). 13. Cui X, Jing X. Stem cell-based therapeutic potential in female ovarian aging and infertility. J Ovarian Res. 2024 Aug 24;17(1):171. doi: 10.1186/s13048-024-01492-3. PMID: 39182123; PMCID: PMC11344413. Kim HK, Kim TJ. 14. Park HS, Ashour D, Elsharoud A, Chugh RM, Ismail N, El Andaloussi A, Al-Hendy A. Towards Cell free Therapy of Premature Ovarian Insufficiency: Human Bone Marrow Mesenchymal Stem Cells Secretome Enhances Angiogenesis in Human Ovarian Microvascular Endothelial Cells. HSOA J Stem Cells Res Dev Ther. 2019;5(2):019. 15. Golshan Sheikhansari, Leili Aghebati-Maleki, Mohammad Nouri, Farhad Jadidi-Niaragh, Mehdi Yousefi. Current approaches for the treatment of premature ovarian failure with stem cell therapy, Biomedicine & Pharmacotherapy, volume 102, 2018,254-262, 16. Sameni HR, Seiri M, Safari M, Tabrizi Amjad MH, Khanmohammadi N, Zarbakhsh S. Bone Marrow Stromal Cells with the Granulocyte Colony-Stimulating Factor in the Management of Chemotherapy-Induced Ovarian Failure in a Rat Model. Iran J Med Sci. 2019 Mar;44(2):135-145. 17. Lee, H.J.; Selesniemi, K.; Niikura, Y.; Niikura, T.; Klein, R.; Dombkowski, D.M.; Tilly, J.L. conducted a study on bone marrow transplantation, which was shown to generate immature oocytes and restore long-term fertility in a preclinical mouse model experiencing chemotherapy-induced premature ovarian failure. Journal of Clinical Oncology , 2007 , 25 , 3198–3204. 18. Liu J, Zhang H, Zhang Y, Li N, Wen Y, Cao F, et al. Homing and restorative effects of bone marrow-derived mesenchymal stem cells on cisplatin-injured ovaries in rats. Mol Cells. 2014;37(12):865. 19. Bao R, et al. Bone marrow derived mesenchymal stem cells transplantation rescues premature ovarian insufficiency induced by chemotherapy. Gynecol Endocrinol. 2018;34(4):320-326. 20. Jane L. Liesveld, Naman Sharma, Omar S. Aljitawi, Stem cell homing: From physiology to therapeutics, Stem Cells , Volume 38, Issue 10, October 2020, Pages 1241–1253 21. Perlin JR, Sporrij A, Zon LI. Blood on the tracks: hematopoietic stem cell-endothelial cell interactions in homing and engraftment. J Mol Med (Berl). 2017 Aug;95(8):809-819. 22. Lifang Yuan, Weiyu Huang, Yin Bi, Saiqiong Chen, Xi Wang, Ting Li, Peiru Wei, Jiebing Du, Ling Zhao, Bo Liu, Yihua Yang, G-CSF-mobilized peripheral blood mononuclear cells combined with platelet-rich plasma restored the ovarian function of aged rats, Journal of Reproductive Immunology,158, 2023. 23. Herraiz S, Romeu M, Buigues A, Martínez S, Díaz-García C, Gómez-Seguí I, et al. Autologous stem cell ovarian transplantation to increase reproductive potential in patients who are poor responders. Fertil Steril. 2018;110(3):496-505. 24. Buigues A, Marchante, M, Miguel-Gómez, L, Martinez, J, Cervelló, I,Pellicer A, HerraizS. Stem cell–secreted factor therapy regenerates the ovarian niche and rescues follicles, American Journal of Obstetrics and Gynecology,Volume 225, Issue 1,2021, Pages 65.e1-65.e14. 25. Buigues A, Ramírez-Martin N, Martínez J, Pellicer N, Meseguer M, Pellicer A, Herraiz S. Systemic changes induced by autologous stem cell ovarian transplant in plasma proteome of women with impaired ovarian reserves. Aging (Albany NY). 2023 Dec 26;15(24):14553-14573. 26. Buigues,A, N Pellicer, J Martínez, N Ramírez-Martín, C Rodríguez-Hernández, P Blázquez, A Pellicer, S Herraiz, P-791 The 4-step ASCOT technique modifies the plasma proteome of patients with premature ovarian insufficiency to resemble that of normoresponder women, Human Reproduction , Volume 39, Issue Supplement_1, July 2024, deae108.1103. Table/Figure Caption List Figure 1 . Schedule for the SC Regenera Treatment. Figure 2. Flow Chart depicting the participant selection, inclusion criteria, and outcomes in the StemCell Regenera Study. Table 1. Characteristic and Demographic data depending on Oocyte activation. Table 2. Outcomes of Stem cell Regenera in OA and Gestational Rate and Leucocytes and CD34+ on fifth day. * P<0.00 ** P<0.5 *** P<0.05 Table 3. Safety of the mobilization in terms of side effects associated with the treatment. Supplementary Material File (figure 1 stemcell regenra.docx) Download 77.67 KB File (figure 2.scregenra.docx) Download 37.52 KB File (table 1.stem cell regenera.docx) Download 11.55 KB File (table 2 stemcellregenera.docx) Download 11.85 KB File (table 3. stemcellregenera.docx) Download 11.43 KB Information & Authors Information Version history V1 Version 1 19 February 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords fertility and assisted reproduction infertility: assisted conception reproductive science: fertilisation reproductive science: ovarian function Authors Affiliations Santamaria 0000-0001-7727-880X [email protected] IVIRMA Alicante Clinic View all articles by this author Ballester European University Department View all articles by this author Muñoz M. IVIRMA Alicante Clinic View all articles by this author Metrics & Citations Metrics Article Usage 453 views 159 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Santamaria, Ballester, Muñoz M.. Effectiveness and Safety of Stem Cell Regeneration for Oocyte Activation in Women with Ovarian Failure: First study using G-CSF mobilization of Peripheral Blood Stem Cells and intraovarian injection of Stem Cell Factor-enriched Platelet Rich Plasma in Clinical Practice. Authorea . 19 February 2025. DOI: https://doi.org/10.22541/au.173994868.86444234/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . 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