Sadeghian-Nodoushan Fatemeh, Aflatoonian Reza, Borzouie Zahra, Akyash Fatemeh, Fesahat Farzaneh, Soleimani Mehrdad, Aghajanpour Samaneh, Moore Harry D, Aflatoonian Behrouz
Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Mol Reprod Dev. 2016 Apr;83(4):312-23. doi: 10.1002/mrd.22620. Epub 2016 Feb 22.
Human male germ-line stem cells (hmGSCs) and human testis-derived embryonic stem cell-like (htESC-like) cells are claimed to be in vitro pluripotent counterparts of spermatogonial stem cells (SSCs), but the origin and pluripotency of human testis-derived cell cultures are still under debate. The aim of this study was to generate putative pluripotent stem cells in vitro from human testicular sperm-extracted (TESE) samples of infertile men, and to assess their pluripotency and capacity to differentiate. TESE samples were minced, enzymatically disaggregated and dispersed into single-cell or cluster suspensions, and then cultured. Initially, cell clusters resembled those described for hmGSCs and htESC-like cells, and were positive for markers such as OCT4/POU5F1, NANOG, and TRA-2-54. Prolonged propagation of cell clusters expressing pluripotency markers did not thrive; instead, the cells that emerged possessed characteristics of mesenchymal stromal cells (MSCs) such as STRO-1, CD105/EGLN1, CD13/ANPEP, SOX9, vimentin, and fibronectin. KIT, SOX2, and CD44 were not expressed by these MSCs. The multipotential differentiation capacity of these cells was confirmed using Oil Red-O and Alizarin Red staining after induction with specific culture conditions. It is therefore concluded that pluripotent stem cells could not be derived using the conditions previously reported to be successful for TESE samples.
人类雄性生殖系干细胞（hmGSCs）和人类睾丸来源的胚胎干细胞样（htESC-like）细胞据称是精原干细胞（SSCs）的体外多能对应物，但人类睾丸来源细胞培养物的起源和多能性仍存在争议。本研究的目的是从不育男性的人类睾丸精子提取（TESE）样本中体外生成假定的多能干细胞，并评估其多能性和分化能力。将TESE样本切碎，酶解并分散成单细胞或细胞团悬液，然后进行培养。最初，细胞团类似于hmGSCs和htESC-like细胞所描述的细胞团，并对OCT4/POU5F1、NANOG和TRA-2-54等标志物呈阳性。表达多能性标志物的细胞团的长期传代培养并不成功；相反，出现的细胞具有间充质基质细胞（MSCs）的特征，如STRO-1、CD105/EGLN1、CD13/ANPEP、SOX9、波形蛋白和纤连蛋白。这些MSCs不表达KIT、SOX2和CD44。在用特定培养条件诱导后，通过油红O和茜素红染色证实了这些细胞的多能分化能力。因此得出结论，使用先前报道对TESE样本成功适用的条件无法获得多能干细胞。
