Document Type : Original Article
Authors
1
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
2
Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
3
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd,Iran.
4
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
5
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi university of Medical Sciences, Yazd, Iran.
10.22074/cellj.2024.2012768.1419
Abstract
Objective: There are ethical and technical challenges in studying human germ cell development. Therefore, in vitro differentiation of human embryonic stem cells (hESCs) as pluripotent cells to the germ cells is a valuable tool to study molecular and cellular aspects of gametogenesis and understanding the causes of infertility.
Methods and Results: Two different complete media (DMEM+20%FBS and EB medium; KOSR-HES without bFGF) were used in both test groups using TCCM and control groups spontaneously differentiated (SD). Thereby, embryoid bodies (EBs) from hESCs (Yazd2; 46XY) were cultured in different conditions including: (i) EB medium (SD-EB), (ii) 60% EB medium+40% conditioned EB medium (TCCM-EB). (iii) 60%EB medium+40% (DMEM+20%FBS) without conditioning (SD-DMEM), (iv) 60%EB medium+40% conditioned DMEM+20%FBS medium (TCCM-DMEM). EBs were collected after 4, 7, and 14 days and their gene expression profiles were assessed and compared with hESCs as day 0 using IF and RT-Q-PCR.
significant increase in the gametogenesis gene expression level in TCCM groups compared with SD groups was determined. Also, immunostaining of differentiated cells in all groups confirmed IVG.
conclusion: Our findings reaveled that human TCCM could be used as a natural niche for in vitro male and female germ cell development.
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