{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:42:34Z","timestamp":1792788154950,"version":"4.5.0"},"reference-count":48,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,5,31]],"date-time":"2025-05-31T00:00:00Z","timestamp":1748649600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,5,31]],"date-time":"2025-05-31T00:00:00Z","timestamp":1748649600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82301794"],"award-info":[{"award-number":["82301794"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Key Research and Development Program of China","award":["2022YFC2702700"],"award-info":[{"award-number":["2022YFC2702700"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Reprod Biol Endocrinol"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Testicular somatic cells play an important role in supporting spermatogenesis. Leydig cells (LCs) and peritubular myoid cells (PTMs) originate from a common progenitor population and show similar expression signatures in adulthood, making it difficult to distinguish and isolate the two in vitro.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In this study, new surface markers for identifying adult LCs (ALCs) and PTMs were discovered by reanalyzing testicular single-cell dataset. Differential expressions of ITGA9 and NGFR were confirmed through immunofluorescence staining of human testes. A novel Fluorescence activated Cell Sorting (FACS) protocol is established for the isolation of ALCs and PTMs based on the two markers. Long-term culture of both cells were performed and their characteristics were characterized and explored.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>ITGA9+ \/NGFR\u2009+\u2009cells were positive for markers of PTMs (SMA, CNN1) and negative for markers of ALCs (HSD3B, STAR), and were able to form tubular and spheroid structures in vitro. In contrast, ITGA9-\/NGFR\u2009+\u2009cells were positive for ALC markers and negative for PTM markers, and showed a capacity of testosterone production in vitro. Also, both cells were negative for Sertoli cell marker SOX9. When the two cells were cultured, they can expand for more than 15 passages.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Our study established a novel and efficient method for identifying and isolating human ALCs and PTMs, which provides a great potential for researches of the two cell types in human.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12958-025-01389-w","type":"journal-article","created":{"date-parts":[[2025,5,31]],"date-time":"2025-05-31T03:19:40Z","timestamp":1748661580000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Identification and isolation of human testicular peritubular myoid cells and Leydig cells by a combination of ITGA9 and NGFR"],"prefix":"10.1186","volume":"23","author":[{"given":"Sha","family":"Han","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangyu","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaqiang","family":"Luo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenkun","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chencheng","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiyong","family":"Ji","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junwei","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruhui","family":"Tian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erlei","family":"Zhi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhua","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuchuan","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,5,31]]},"reference":[{"issue":"1","key":"1389_CR1","doi-asserted-by":"publisher","first-page":"5683","DOI":"10.1038\/s41467-020-19414-4","volume":"11","author":"L Zhao","year":"2020","unstructured":"Zhao L, et al. Single-cell analysis of developing and azoospermia human testicles reveals central role of Sertoli cells. Nat Commun. 2020;11(1):5683.","journal-title":"Nat Commun"},{"issue":"6","key":"1389_CR2","doi-asserted-by":"publisher","first-page":"1589","DOI":"10.1016\/j.celrep.2018.01.043","volume":"22","author":"I Stevant","year":"2018","unstructured":"Stevant I, et al. Deciphering cell lineage specification during male sex determination with Single-Cell RNA sequencing. Cell Rep. 2018;22(6):1589\u201399.","journal-title":"Cell Rep"},{"issue":"2","key":"1389_CR3","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1016\/j.stem.2019.12.005","volume":"26","author":"J Guo","year":"2020","unstructured":"Guo J, et al. The dynamic transcriptional cell atlas of testis development during human puberty. Cell Stem Cell. 2020;26(2):262\u2013e2764.","journal-title":"Cell Stem Cell"},{"key":"1389_CR4","doi-asserted-by":"publisher","first-page":"111179","DOI":"10.1016\/j.mce.2021.111179","volume":"525","author":"X Zhao","year":"2021","unstructured":"Zhao X, et al. Differentiation of seminiferous tubule-associated stem cells into Leydig cell and myoid cell lineages. Mol Cell Endocrinol. 2021;525:111179.","journal-title":"Mol Cell Endocrinol"},{"issue":"12","key":"1389_CR5","doi-asserted-by":"publisher","first-page":"2663","DOI":"10.1093\/humrep\/deaa271","volume":"35","author":"V Mularoni","year":"2020","unstructured":"Mularoni V, et al. Age-related changes in human Leydig cell status. Hum Reprod. 2020;35(12):2663\u201376.","journal-title":"Hum Reprod"},{"issue":"10","key":"1389_CR6","doi-asserted-by":"publisher","first-page":"e3123","DOI":"10.1038\/cddis.2017.531","volume":"8","author":"M Zhang","year":"2017","unstructured":"Zhang M, et al. Transplanted human p75-positive stem Leydig cells replace disrupted Leydig cells for testosterone production. Cell Death Dis. 2017;8(10):e3123.","journal-title":"Cell Death Dis"},{"issue":"9","key":"1389_CR7","doi-asserted-by":"publisher","first-page":"1621","DOI":"10.1093\/humrep\/dez131","volume":"34","author":"J Eliveld","year":"2019","unstructured":"Eliveld J, et al. Primary human testicular PDGFRalpha\u2009+\u2009cells are multipotent and can be differentiated into cells with Leydig cell characteristics in vitro. Hum Reprod. 2019;34(9):1621\u201331.","journal-title":"Hum Reprod"},{"issue":"10","key":"1389_CR8","doi-asserted-by":"publisher","first-page":"2197","DOI":"10.1093\/humrep\/deaa174","volume":"35","author":"K Xia","year":"2020","unstructured":"Xia K, et al. Endosialin defines human stem Leydig cells with regenerative potential. Hum Reprod. 2020;35(10):2197\u2013212.","journal-title":"Hum Reprod"},{"issue":"5","key":"1389_CR9","doi-asserted-by":"publisher","first-page":"1265","DOI":"10.1111\/andr.12817","volume":"8","author":"J Eliveld","year":"2020","unstructured":"Eliveld J, et al. A comparative analysis of human adult testicular cells expressing stem Leydig cell markers in the interstitium, vasculature, and peritubular layer. Andrology. 2020;8(5):1265\u201376.","journal-title":"Andrology"},{"issue":"2","key":"1389_CR10","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1530\/REP-17-0346","volume":"155","author":"M Gaur","year":"2018","unstructured":"Gaur M, et al. Isolation of human testicular cells and co-culture with embryonic stem cells. Reproduction. 2018;155(2):153\u201366.","journal-title":"Reproduction"},{"issue":"4","key":"1389_CR11","doi-asserted-by":"publisher","first-page":"898","DOI":"10.1111\/andr.12669","volume":"8","author":"A Mayerhofer","year":"2020","unstructured":"Mayerhofer A. Peritubular cells of the human testis: prostaglandin E2 and more. Andrology. 2020;8(4):898\u2013902.","journal-title":"Andrology"},{"issue":"5","key":"1389_CR12","doi-asserted-by":"publisher","first-page":"R107","DOI":"10.1530\/REP-12-0497","volume":"145","author":"A Mayerhofer","year":"2013","unstructured":"Mayerhofer A. Human testicular peritubular cells: more than Meets the eye. Reproduction. 2013;145(5):R107\u201316.","journal-title":"Reproduction"},{"issue":"5","key":"1389_CR13","doi-asserted-by":"publisher","first-page":"1956","DOI":"10.1210\/jc.2005-2169","volume":"91","author":"M Albrecht","year":"2006","unstructured":"Albrecht M, et al. Isolation and cultivation of human testicular peritubular cells: a new model for the investigation of fibrotic processes in the human testis and male infertility. J Clin Endocrinol Metab. 2006;91(5):1956\u201360.","journal-title":"J Clin Endocrinol Metab"},{"key":"1389_CR14","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1016\/j.mce.2018.01.022","volume":"473","author":"V Rey-Ares","year":"2018","unstructured":"Rey-Ares V, et al. Prostaglandin E2 (PGE2) is a testicular peritubular cell-derived factor involved in human testicular homeostasis. Mol Cell Endocrinol. 2018;473:217\u201324.","journal-title":"Mol Cell Endocrinol"},{"key":"1389_CR15","doi-asserted-by":"publisher","first-page":"11945","DOI":"10.1038\/ncomms11945","volume":"7","author":"RC Rennert","year":"2016","unstructured":"Rennert RC, et al. Microfluidic single-cell transcriptional analysis rationally identifies novel surface marker profiles to enhance cell-based therapies. Nat Commun. 2016;7:11945.","journal-title":"Nat Commun"},{"issue":"23","key":"1389_CR16","doi-asserted-by":"publisher","first-page":"5429","DOI":"10.1182\/blood-2011-11-393645","volume":"119","author":"A Nakamura-Ishizu","year":"2012","unstructured":"Nakamura-Ishizu A, et al. Extracellular matrix protein tenascin-C is required in the bone marrow microenvironment primed for hematopoietic regeneration. Blood. 2012;119(23):5429\u201337.","journal-title":"Blood"},{"issue":"1\u20132","key":"1389_CR17","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1007\/s11010-014-2212-2","volume":"398","author":"J Zhang","year":"2015","unstructured":"Zhang J, et al. Effects of p75 neurotrophin receptor on regulating hypoxia-induced angiogenic factors in retinal pigment epithelial cells. Mol Cell Biochem. 2015;398(1\u20132):123\u201334.","journal-title":"Mol Cell Biochem"},{"issue":"5","key":"1389_CR18","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1038\/s43587-024-00614-2","volume":"4","author":"Z Deng","year":"2024","unstructured":"Deng Z, et al. Targeting dysregulated phago-\/auto-lysosomes in Sertoli cells to ameliorate late-onset hypogonadism. Nat Aging. 2024;4(5):647\u201363.","journal-title":"Nat Aging"},{"issue":"1","key":"1389_CR19","doi-asserted-by":"publisher","first-page":"7494","DOI":"10.1038\/s41467-022-34460-w","volume":"13","author":"Y Qu","year":"2022","unstructured":"Qu Y, et al. Proteogenomic characterization of MiT family translocation renal cell carcinoma. Nat Commun. 2022;13(1):7494.","journal-title":"Nat Commun"},{"issue":"3","key":"1389_CR20","doi-asserted-by":"publisher","first-page":"1562","DOI":"10.1007\/s12035-023-03633-z","volume":"61","author":"M Jung","year":"2024","unstructured":"Jung M, et al. Neuronal stem cells from Late-Onset alzheimer patients show altered regulation of Sirtuin 1 depending on Apolipoprotein E indicating disturbed stem cell plasticity. Mol Neurobiol. 2024;61(3):1562\u201379.","journal-title":"Mol Neurobiol"},{"issue":"1","key":"1389_CR21","doi-asserted-by":"publisher","first-page":"3164","DOI":"10.1038\/s41467-019-11036-9","volume":"10","author":"X Fan","year":"2019","unstructured":"Fan X, et al. Single-cell reconstruction of follicular remodeling in the human adult ovary. Nat Commun. 2019;10(1):3164.","journal-title":"Nat Commun"},{"key":"1389_CR22","doi-asserted-by":"crossref","unstructured":"Han S et al. Low-Intensity pulsed ultrasound alleviates human testicular Leydig cell senescence in vitro. Int J Mol Sci, 2022. 24(1).","DOI":"10.3390\/ijms24010418"},{"issue":"6","key":"1389_CR23","doi-asserted-by":"publisher","first-page":"1648","DOI":"10.1093\/biolre\/ioz053","volume":"100","author":"S Sakib","year":"2019","unstructured":"Sakib S, et al. Formation of organotypic testicular organoids in microwell Culturedagger. Biol Reprod. 2019;100(6):1648\u201360.","journal-title":"Biol Reprod"},{"issue":"2","key":"1389_CR24","doi-asserted-by":"publisher","first-page":"259","DOI":"10.1530\/REP-20-0100","volume":"160","author":"N Schmid","year":"2020","unstructured":"Schmid N, et al. A translational cellular model for the study of peritubular cells of the testis. Reproduction. 2020;160(2):259\u201368.","journal-title":"Reproduction"},{"issue":"5","key":"1389_CR25","doi-asserted-by":"publisher","first-page":"756","DOI":"10.1111\/andr.12509","volume":"6","author":"C Mayer","year":"2018","unstructured":"Mayer C, et al. Insights into the role of androgen receptor in human testicular peritubular cells. Andrology. 2018;6(5):756\u201365.","journal-title":"Andrology"},{"issue":"1","key":"1389_CR26","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1111\/j.1365-2605.1993.tb01149.x","volume":"16","author":"H Lejeune","year":"1993","unstructured":"Lejeune H, et al. Enhancement of testosterone secretion by normal adult human Leydig cells by co-culture with enriched preparations of normal adult human Sertoli cells. Int J Androl. 1993;16(1):27\u201334.","journal-title":"Int J Androl"},{"issue":"5","key":"1389_CR27","first-page":"1426","volume":"82","author":"C Cudicini","year":"1997","unstructured":"Cudicini C, et al. Human Leydig cells and Sertoli cells are producers of interleukins-1 and\u2013\u20096. J Clin Endocrinol Metab. 1997;82(5):1426\u201333.","journal-title":"J Clin Endocrinol Metab"},{"issue":"3","key":"1389_CR28","first-page":"129","volume":"21","author":"H Lejeune","year":"1998","unstructured":"Lejeune H, Sanchez P, Saez JM. Enhancement of long-term testosterone secretion and steroidogenic enzyme expression in human Leydig cells by co-culture with human Sertoli cell-enriched preparations. Int J Androl. 1998;21(3):129\u201340.","journal-title":"Int J Androl"},{"issue":"1","key":"1389_CR29","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1002\/jcp.20805","volume":"210","author":"MH Perrard","year":"2007","unstructured":"Perrard MH, et al. beta-Nerve growth factor participates in an auto\/paracrine pathway of regulation of the meiotic differentiation of rat spermatocytes. J Cell Physiol. 2007;210(1):51\u201362.","journal-title":"J Cell Physiol"},{"issue":"10","key":"1389_CR30","doi-asserted-by":"publisher","first-page":"4755","DOI":"10.1210\/en.2009-0416","volume":"150","author":"P Lim","year":"2009","unstructured":"Lim P, et al. Sertoli cell androgen receptor DNA binding domain is essential for the completion of spermatogenesis. Endocrinology. 2009;150(10):4755\u201365.","journal-title":"Endocrinology"},{"issue":"2","key":"1389_CR31","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1016\/j.bbrc.2013.05.098","volume":"436","author":"L Zhang","year":"2013","unstructured":"Zhang L, et al. NGF induces adult stem Leydig cells to proliferate and differentiate during Leydig cell regeneration. Biochem Biophys Res Commun. 2013;436(2):300\u20135.","journal-title":"Biochem Biophys Res Commun"},{"issue":"1","key":"1389_CR32","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1385\/ENDO:29:1:155","volume":"29","author":"W Jin","year":"2006","unstructured":"Jin W, et al. Cellular localization of NGF and its receptors TrkA and p75LNGFR in male reproductive organs of the Japanese monkey, Macaca fuscata fuscata. Endocrine. 2006;29(1):155\u201360.","journal-title":"Endocrine"},{"issue":"3","key":"1389_CR33","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1111\/j.1469-7580.2006.00528.x","volume":"208","author":"MB Levanti","year":"2006","unstructured":"Levanti MB, et al. Effects of increased nerve growth factor plasma levels on the expression of TrkA and p75 in rat testicles. J Anat. 2006;208(3):373\u20139.","journal-title":"J Anat"},{"issue":"9","key":"1389_CR34","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.1111\/and.12375","volume":"47","author":"K Lin","year":"2015","unstructured":"Lin K, et al. Nerve growth factor promotes human sperm motility in vitro by increasing the movement distance and the number of A grade spermatozoa. Andrologia. 2015;47(9):1041\u20136.","journal-title":"Andrologia"},{"issue":"6","key":"1389_CR35","doi-asserted-by":"publisher","first-page":"771","DOI":"10.1016\/j.beem.2012.05.002","volume":"26","author":"V Papadopoulos","year":"2012","unstructured":"Papadopoulos V, Miller WL. Role of mitochondria in steroidogenesis. Best Pract Res Clin Endocrinol Metab. 2012;26(6):771\u201390.","journal-title":"Best Pract Res Clin Endocrinol Metab"},{"issue":"11","key":"1389_CR36","doi-asserted-by":"publisher","first-page":"1868","DOI":"10.1210\/me.2012-1159","volume":"26","author":"MB Rone","year":"2012","unstructured":"Rone MB, et al. Identification of a dynamic mitochondrial protein complex driving cholesterol import, trafficking, and metabolism to steroid hormones. Mol Endocrinol. 2012;26(11):1868\u201382.","journal-title":"Mol Endocrinol"},{"key":"1389_CR37","doi-asserted-by":"crossref","unstructured":"Stockl JB et al. Proteomic insights into senescence of testicular peritubular cells from a nonhuman primate model. Cells, 2020. 9(11).","DOI":"10.3390\/cells9112498"},{"issue":"9","key":"1389_CR38","doi-asserted-by":"publisher","first-page":"1160","DOI":"10.1016\/j.devcel.2022.04.004","volume":"57","author":"X Nie","year":"2022","unstructured":"Nie X, et al. Single-cell analysis of human testis aging and correlation with elevated body mass index. Dev Cell. 2022;57(9):1160\u2013e11765.","journal-title":"Dev Cell"},{"key":"1389_CR39","doi-asserted-by":"crossref","unstructured":"DeCicco-Skinner KL et al. Endothelial cell tube formation assay for the in vitro study of angiogenesis. J Vis Exp, 2014(91): p. e51312.","DOI":"10.3791\/51312-v"},{"key":"1389_CR40","doi-asserted-by":"crossref","unstructured":"Stryker ZI et al. Evaluation Angiogenesis Assays Biomedicines, 2019. 7(2).","DOI":"10.3390\/biomedicines7020037"},{"issue":"4","key":"1389_CR41","doi-asserted-by":"publisher","first-page":"764","DOI":"10.1016\/j.stem.2020.12.004","volume":"28","author":"J Guo","year":"2021","unstructured":"Guo J, et al. Single-cell analysis of the developing human testis reveals somatic niche cell specification and fetal germline stem cell establishment. Cell Stem Cell. 2021;28(4):764\u2013e7784.","journal-title":"Cell Stem Cell"},{"issue":"4","key":"1389_CR42","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1007\/s10456-020-09732-y","volume":"23","author":"S Del Diaz","year":"2020","unstructured":"Del Diaz S, et al. Embryonic Circulating endothelial progenitor cells. Angiogenesis. 2020;23(4):531\u201341.","journal-title":"Angiogenesis"},{"issue":"2","key":"1389_CR43","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1093\/molehr\/gax063","volume":"24","author":"M Mincheva","year":"2018","unstructured":"Mincheva M, et al. Reassembly of adult human testicular cells: can testis cord-like structures be created in vitro? Mol Hum Reprod. 2018;24(2):55\u201363.","journal-title":"Mol Hum Reprod"},{"issue":"2","key":"1389_CR44","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1038\/nprot.2017.140","volume":"13","author":"JP Alves-Lopes","year":"2018","unstructured":"Alves-Lopes JP, Soder O, Stukenborg JB. Use of a three-layer gradient system of cells for rat testicular organoid generation. Nat Protoc. 2018;13(2):248\u201359.","journal-title":"Nat Protoc"},{"issue":"16","key":"1389_CR45","doi-asserted-by":"publisher","first-page":"4492","DOI":"10.1016\/j.biomaterials.2010.02.029","volume":"31","author":"A Legendre","year":"2010","unstructured":"Legendre A, et al. An engineered 3D blood-testis barrier model for the assessment of reproductive toxicity potential. Biomaterials. 2010;31(16):4492\u2013505.","journal-title":"Biomaterials"},{"key":"1389_CR46","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.addr.2014.01.003","volume":"74","author":"M Matsusaki","year":"2014","unstructured":"Matsusaki M, Case CP, Akashi M. Three-dimensional cell culture technique and pathophysiology. Adv Drug Deliv Rev. 2014;74:95\u2013103.","journal-title":"Adv Drug Deliv Rev"},{"issue":"7","key":"1389_CR47","doi-asserted-by":"publisher","first-page":"879","DOI":"10.1038\/nbt1109","volume":"23","author":"S Levenberg","year":"2005","unstructured":"Levenberg S, et al. Engineering vascularized skeletal muscle tissue. Nat Biotechnol. 2005;23(7):879\u201384.","journal-title":"Nat Biotechnol"},{"issue":"7","key":"1389_CR48","doi-asserted-by":"publisher","first-page":"1646","DOI":"10.1016\/j.biomaterials.2009.11.036","volume":"31","author":"T Sasagawa","year":"2010","unstructured":"Sasagawa T, et al. Design of prevascularized three-dimensional cell-dense tissues using a cell sheet stacking manipulation technology. Biomaterials. 2010;31(7):1646\u201354.","journal-title":"Biomaterials"}],"container-title":["Reproductive Biology and Endocrinology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12958-025-01389-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12958-025-01389-w","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12958-025-01389-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T10:40:51Z","timestamp":1781606451000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s12958-025-01389-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,31]]},"references-count":48,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["1389"],"URL":"https:\/\/doi.org\/10.1186\/s12958-025-01389-w","relation":{},"ISSN":["1477-7827"],"issn-type":[{"value":"1477-7827","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,31]]},"assertion":[{"value":"11 October 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"29 March 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 May 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This study was approved by the Ethical Review Committee of Shanghai General Hospital on 2022-3-22 (Approval number:2022SQ294).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"82"}}