title: Pluripotency and differentiation of cells from human testicular sperm extraction: An investigation of cell stemness.
abstract: 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.
authors.affiliation: Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Sadeghian-Nodoushan
authors.affiliation: Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Aflatoonian
authors.affiliation: Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Borzouie
authors.affiliation: Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Akyash
authors.affiliation: Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Fesahat
authors.affiliation: Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Samaneh
authors.initials: S
authors.lastname: Aghajanpour
authors.affiliation: Centre for Stem Cell Biology (CSCB), Department of Biomedical Sciences, The University of Sheffield, Western Bank, Alfred Denny Building, Sheffield, United Kingdom.
authors.forename: Harry D
authors.initials: HD
authors.lastname: Moore
authors.affiliation: Stem Cell Biology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Behrouz
authors.initials: B
authors.lastname: Aflatoonian
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D020869:Gene Expression Profiling; D006801:Humans; D007248:Infertility, Male; D008297:Male; D039904:Pluripotent Stem Cells / Q000166:cytology*; D053625:Sperm Retrieval; D013091:Spermatogenesis / Q000235:genetics; D013093:Spermatogonia / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology; D013737:Testis / Q000166:cytology*