{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T07:31:21Z","timestamp":1792481481815,"version":"4.5.0"},"reference-count":47,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2022,8,20]],"date-time":"2022-08-20T00:00:00Z","timestamp":1660953600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the State Assignment of Lomonosov MSU"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Idiopathic male infertility is a highly prevalent diagnosis in developed countries with no specific treatment options. Although empirical medical treatment is widely used to restore male fertility, its efficacy remains limited and inconclusively proven. Therefore, the development of novel therapeutic approaches in this field is a high-priority task. Since the failure of testicular microenvironment components might be involved in the pathogenesis of idiopathic male infertility, application of mesenchymal stromal cells (MSCs) as well as the MSC secretome is worth considering. Previously, we showed that the intratesticular injection of MSCs or the MSC secretome led to the recovery of spermatogenesis at least through replenishing the testicular microenvironment and its maintenance by MSC-secreted paracrine factors. However, the clinical use of such products has been limited to single trials to date. This may be due to the lack of relevant potency tests reflecting mechanisms of action of the MSC secretome in male infertility models. Based on the presumptive MSC secretome mode of action on the testicular microenvironment, we suggest a novel approach to test the potential efficacy of the MSC secretome for idiopathic male infertility treatment. It represents a potency assay based on evaluation of testosterone production by isolated Leydig cells. We demonstrated that the MSC secretome stimulated testosterone secretion by Leydig cells in vitro. We then hypothesized that among the major factors of the MSC secretome, vascular endothelial growth factor (VEGF) could be responsible for the observed effects, which we confirmed by the revealed correlation between the extent of stimulated testosterone production and VEGF concentration in the MSC secretome. The pilot results obtained from the doxorubicin-induced male infertility murine model also indicate the important impact of VEGF in the MSC secretome\u2019s regenerative effects. Utilizing VEGF as a surrogate factor, a novel approach to study the potency of MSC secretome-based products for idiopathic male infertility treatment is suggested. Further validation is required for its implementation into the biopharmaceutical manufacturing process.<\/jats:p>","DOI":"10.3390\/ijms23169414","type":"journal-article","created":{"date-parts":[[2022,8,21]],"date-time":"2022-08-21T22:23:13Z","timestamp":1661120593000},"page":"9414","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Novel Potency Assay for MSC Secretome-Based Treatment of Idiopathic Male Infertility Employed Leydig Cells and Revealed Vascular Endothelial Growth Factor as a Promising Potency Marker"],"prefix":"10.3390","volume":"23","author":[{"given":"Anna","family":"Monakova","sequence":"first","affiliation":[{"name":"Faculty of Medicine, Lomonosov Moscow State University, 27\/1, Lomonosovskiy Av., Moscow 119192, Russia"},{"name":"Institute for Regenerative Medicine, Medical Research and Education Center, Lomonosov Moscow State University, 27\/10, Lomonosovskiy Av., Moscow 119192, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georgy","family":"Sagaradze","sequence":"additional","affiliation":[{"name":"Institute for Regenerative Medicine, Medical Research and Education Center, Lomonosov Moscow State University, 27\/10, Lomonosovskiy Av., Moscow 119192, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2597-8879","authenticated-orcid":false,"given":"Nataliya","family":"Basalova","sequence":"additional","affiliation":[{"name":"Institute for Regenerative Medicine, Medical Research and Education Center, Lomonosov Moscow State University, 27\/10, Lomonosovskiy Av., Moscow 119192, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5039-7152","authenticated-orcid":false,"given":"Vladimir","family":"Popov","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, Lomonosov Moscow State University, 27\/1, Lomonosovskiy Av., Moscow 119192, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vadim","family":"Balabanyan","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, Lomonosov Moscow State University, 27\/1, Lomonosovskiy Av., Moscow 119192, Russia"},{"name":"Institute for Regenerative Medicine, Medical Research and Education Center, Lomonosov Moscow State University, 27\/10, Lomonosovskiy Av., Moscow 119192, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0696-1369","authenticated-orcid":false,"given":"Anastasia","family":"Efimenko","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, Lomonosov Moscow State University, 27\/1, Lomonosovskiy Av., Moscow 119192, Russia"},{"name":"Institute for Regenerative Medicine, Medical Research and Education Center, Lomonosov Moscow State University, 27\/10, Lomonosovskiy Av., Moscow 119192, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/S0140-6736(20)32667-2","article-title":"Male Infertility","volume":"397","author":"Agarwal","year":"2021","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"638628","DOI":"10.3389\/fphar.2021.638628","article-title":"Effectiveness of Pharmacological Intervention Among Men with Infertility: A Systematic Review and Network Meta-Analysis","volume":"12","author":"Shahid","year":"2021","journal-title":"Front. Pharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1517","DOI":"10.1080\/14656566.2019.1615057","article-title":"Effects of the Selective Estrogen Receptor Modulators for the Treatment of Male Infertility: A Systematic Review and Meta-Analysis","volume":"20","author":"Cannarella","year":"2019","journal-title":"Expert Opin. Pharmacother."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1016\/j.fertnstert.2019.11.008","article-title":"The Effect of Antioxidants on Male Factor Infertility: The Males, Antioxidants, and Infertility (MOXI) Randomized Clinical Trial","volume":"113","author":"Steiner","year":"2020","journal-title":"Fertil. Steril."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.bj.2022.01.015","article-title":"Regenerative Medicine for Male Infertility: A Focus on Stem Cell Niche Injury Models","volume":"45","author":"Sagaradze","year":"2022","journal-title":"Biomed. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1186\/s13287-021-02295-9","article-title":"Mesenchymal Stromal\/stem Cells and Their Exosomes for Restoration of Spermatogenesis in Non-Obstructive Azoospermia: A Systemic Review","volume":"12","author":"Zhankina","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1111\/andr.12713","article-title":"Potential Use of Stem Cells for Fertility Preservation","volume":"8","author":"Kauffman","year":"2020","journal-title":"Andrology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.1016\/j.biopha.2018.10.174","article-title":"Application of Rat Cryptorchidism Model for the Evaluation of Mesenchymal Stromal Cell Secretome Regenerative Potential","volume":"109","author":"Sagaradze","year":"2019","journal-title":"Biomed. Pharmacother."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8450721","DOI":"10.1155\/2021\/8450721","article-title":"Therapeutic Effect of Stem Cells on Male Infertility in a Rat Model: Histological, Molecular, Biochemical, and Functional Study","volume":"2021","author":"Mohammed","year":"2021","journal-title":"Stem Cells Int."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1002\/stem.2847","article-title":"Concise Review: Using Fat to Fight Disease: A Systematic Review of Nonhomologous Adipose-Derived Stromal\/Stem Cell Therapies","volume":"36","author":"Bateman","year":"2018","journal-title":"Stem Cells"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/s12929-021-00725-7","article-title":"Mesenchymal Stem\/stromal Cell-Based Therapy: Mechanism, Systemic Safety and Biodistribution for Precision Clinical Applications","volume":"28","author":"Zhuang","year":"2021","journal-title":"J. Biomed. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1016\/j.jcyt.2019.08.002","article-title":"Mesenchymal Stem versus Stromal Cells: International Society for Cell & Gene Therapy (ISCT\u00ae) Mesenchymal Stromal Cell Committee Position Statement on Nomenclature","volume":"21","author":"Viswanathan","year":"2019","journal-title":"Cytotherapy"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.jcyt.2014.04.002","article-title":"Standardization of Good Manufacturing Practice-Compliant Production of Bone Marrow-Derived Human Mesenchymal Stromal Cells for Immunotherapeutic Applications","volume":"17","author":"Wuchter","year":"2015","journal-title":"Cytotherapy"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"576176","DOI":"10.3389\/fcell.2020.576176","article-title":"Mesenchymal Stromal Cells as Critical Contributors to Tissue Regeneration","volume":"8","author":"Sagaradze","year":"2020","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"661532","DOI":"10.3389\/fcell.2021.661532","article-title":"The Therapeutic Potential of Mesenchymal Stromal Cells for Regenerative Medicine: Current Knowledge and Future Understandings","volume":"9","author":"Merimi","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"753","DOI":"10.2217\/nnm-2018-0240","article-title":"Freeze-Dried and GMP-Compliant Pharmaceuticals Containing Exosomes for Acellular Mesenchymal Stromal Cell Immunomodulant Therapy","volume":"14","author":"Bari","year":"2019","journal-title":"Nanomedicine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.jcyt.2021.01.001","article-title":"Critical Considerations for the Development of Potency Tests for Therapeutic Applications of Mesenchymal Stromal Cell-Derived Small Extracellular Vesicles","volume":"23","author":"Gimona","year":"2021","journal-title":"Cytotherapy"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1016\/j.jcyt.2017.03.076","article-title":"Regulatory Perspective on in Vitro Potency Assays for Human Mesenchymal Stromal Cells Used in Immunotherapy","volume":"19","author":"Hoefnagel","year":"2017","journal-title":"Cytotherapy"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1186\/s13287-017-0488-3","article-title":"Development of a Surrogate Potency Assay to Determine the Angiogenic Activity of Stempeucel\u00ae, a Pooled, Ex-Vivo Expanded, Allogeneic Human Bone Marrow Mesenchymal Stromal Cell Product","volume":"8","author":"Thej","year":"2017","journal-title":"Stem Cell Res. Ther."},{"key":"ref_20","unstructured":"European Medicines Agency (1999). ICH Topic Q 6 B Specifications: Test Procedures and Acceptance Criteria for Biotechnological\/Biological Products."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"E1763","DOI":"10.1152\/ajpendo.00611.2006","article-title":"Effect of Hypoxia on the Release of Vascular Endothelial Growth Factor and Testosterone in Mouse TM3 Leydig Cells","volume":"292","author":"Hwang","year":"2007","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1186\/s13287-019-1479-3","article-title":"A Magic Kick for Regeneration: Role of Mesenchymal Stromal Cell Secretome in Spermatogonial Stem Cell Niche Recovery","volume":"10","author":"Sagaradze","year":"2019","journal-title":"Stem Cell Res. Ther."},{"key":"ref_23","first-page":"e53389","article-title":"Isolation of Sertoli Cells and Peritubular Cells from Rat Testes","volume":"108","author":"Bhushan","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_24","first-page":"18","article-title":"Causes of Male Infertility: A 9-Year Prospective Monocentre Study on 1737 Patients with Reduced Total Sperm Counts","volume":"32","author":"Punab","year":"2017","journal-title":"Hum. Reprod."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"810242","DOI":"10.3389\/fendo.2022.810242","article-title":"The Effect of Antioxidants on Sperm Quality Parameters and Pregnancy Rates for Idiopathic Male Infertility: A Network Meta-Analysis of Randomized Controlled Trials","volume":"151","author":"Li","year":"2022","journal-title":"Front. Endocrinol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1945","DOI":"10.1093\/humrep\/16.9.1945","article-title":"Significance of Serum Inhibin B Concentration for Evaluating Improvement in Spermatogenesis after Varicocelectomy","volume":"16","author":"Fujisawa","year":"2001","journal-title":"Hum. Reprod."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"746","DOI":"10.1093\/humrep\/12.4.746","article-title":"Physiological Relationships between Inhibin B, Follicle Stimulating Hormone Secretion and Spermatogenesis in Normal Men and Response to Gonadotrophin Suppression by Exogenous Testosterone","volume":"12","author":"Anderson","year":"1997","journal-title":"Hum. Reprod."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/0303-7207(94)90191-0","article-title":"A Sertoli Cell-Secreted Paracrine Factor(s) Stimulates Proliferation and Inhibits Steroidogenesis of Rat Leydig Cells","volume":"106","author":"Wu","year":"1994","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"R18","DOI":"10.1152\/ajpregu.00327.2006","article-title":"An Integrated View of Oxidative Stress in Aging: Basic Mechanisms, Functional Effects, and Pathological Considerations","volume":"292","author":"Kregel","year":"2007","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1530\/REP-09-0020","article-title":"Vascular Endothelial Growth Factor Regulates Germ Cell Survival during Establishment of Spermatogenesis in the Bovine Testis","volume":"138","author":"Caires","year":"2009","journal-title":"Reproduction"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1095\/biolreprod65.4.1201","article-title":"Intercellular Communication between Sertoli Cells and Leydig Cells in the Absence of Follicle-Stimulating Hormone-Receptor Signaling","volume":"65","author":"Krishnamurthy","year":"2001","journal-title":"Biol. Reprod."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4379","DOI":"10.1038\/s41467-018-06881-z","article-title":"Testicular Endothelial Cells Are a Critical Population in the Germline Stem Cell Niche","volume":"9","author":"Bhang","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12820","DOI":"10.1038\/srep12820","article-title":"Human Testicular Peritubular Cells Secrete Pigment Epithelium-Derived Factor (PEDF), Which May Be Responsible for the Avascularity of the Seminiferous Tubules","volume":"5","author":"Kampfer","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1905","DOI":"10.1007\/s12015-021-10197-9","article-title":"Sertoli Cells Possess Immunomodulatory Properties and the Ability of Mitochondrial Transfer Similar to Mesenchymal Stromal Cells","volume":"17","author":"Porubska","year":"2021","journal-title":"Stem Cell Rev. Rep."},{"key":"ref_35","first-page":"1067","article-title":"Are Sertoli Cells a Kind of Mesenchymal Stem Cells?","volume":"9","author":"Gong","year":"2017","journal-title":"Am. J. Transl. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.jcyt.2015.11.008","article-title":"International Society for Cellular Therapy Perspective on Immune Functional Assays for Mesenchymal Stromal Cells as Potency Release Criterion for Advanced Phase Clinical Trials","volume":"18","author":"Galipeau","year":"2016","journal-title":"Cytotherapy"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"994","DOI":"10.1095\/biolreprod40.5.994","article-title":"Temporal Appearance of Seasonal Changes in Numbers of Sertoli Cells, Leydig Cells, and Germ Cells in Stallions","volume":"40","author":"Johnson","year":"1989","journal-title":"Biol. Reprod."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Shin, S., Lee, J., Kwon, Y., Park, K.-S., Jeong, J.-H., Choi, S.-J., Bang, S.I., Chang, J.W., and Lee, C. (2021). Comparative Proteomic Analysis of the Mesenchymal Stem Cells Secretome from Adipose, Bone Marrow, Placenta and Wharton\u2019s Jelly. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22020845"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2474","DOI":"10.3389\/fimmu.2019.02474","article-title":"Multi-Parameter Analysis of Biobanked Human Bone Marrow Stromal Cells Shows Little Influence for Donor Age and Mild Comorbidities on Phenotypic and Functional Properties","volume":"10","author":"Andrzejewska","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/1479-5876-12-8","article-title":"Donor Age Negatively Impacts Adipose Tissue-Derived Mesenchymal Stem Cell Expansion and Differentiation","volume":"12","author":"Choudhery","year":"2014","journal-title":"J. Transl. Med."},{"key":"ref_41","first-page":"454","article-title":"Adipose Stem Cell Function Maintained with Age: An Intra-Subject Study of Long-Term Cryopreserved Cells","volume":"37","author":"Kokai","year":"2017","journal-title":"Aesthet. Surg. J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1186\/s13287-018-0914-1","article-title":"Adipose-Derived and Bone Marrow Mesenchymal Stem Cells: A Donor-Matched Comparison","volume":"9","author":"Fristad","year":"2018","journal-title":"Stem Cell Res. Ther."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1186\/s13287-018-0876-3","article-title":"Changes in Phenotype and Differentiation Potential of Human Mesenchymal Stem Cells Aging in Vitro","volume":"9","author":"Yang","year":"2018","journal-title":"Stem Cell Res. Ther."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Sagaradze, G., Grigorieva, O., Nimiritsky, P., Basalova, N., Kalinina, N., Akopyan, Z., and Efimenko, A. (2019). Conditioned Medium from Human Mesenchymal Stromal Cells: Towards the Clinical Translation. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20071656"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1186\/s13287-017-0716-x","article-title":"Tissue Source Determines the Differentiation Potentials of Mesenchymal Stem Cells: A Comparative Study of Human Mesenchymal Stem Cells from Bone Marrow and Adipose Tissue","volume":"8","author":"Xu","year":"2017","journal-title":"Stem Cell Res. Ther."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1080\/14653240600855905","article-title":"Minimal Criteria for Defining Multipotent Mesenchymal Stromal Cells. The International Society for Cellular Therapy Position Statement","volume":"8","author":"Dominici","year":"2006","journal-title":"Cytotherapy"},{"key":"ref_47","unstructured":"(2022, August 01). Available online: https:\/\/imagej.net\/contribute\/citing."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/16\/9414\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:12:57Z","timestamp":1760141577000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/16\/9414"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,20]]},"references-count":47,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["ijms23169414"],"URL":"https:\/\/doi.org\/10.3390\/ijms23169414","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,20]]}}}