Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions

Abstract

We found that hSSCs show limited proliferation in vitro under mSSC culture conditions. Coculture of HLA/EPCAM sorted cells with testicular feeders improved the germ cell/somatic cell ratio.

In vitro culture of spermatogonial stem cells isolated from adult alpaca (Vicugna pacos) testes analysed withDolichos biflorusby flow cytometry

“…On the other hand, the proportion of SSCs remained similar between 14 and 21 days, this would indicate that the alpaca cells during in vitro culture period were without change in the rate of SSC that begun the differentiation. Medrano, Rombaut, Simon, Pellicer, and Goossens () observed in human a major SSC proliferation from 7 to 14 days of culture, afterwards SSC decreased dramatically by the day 28; this supports our results, where alpaca SSC during 14 and 21 days was higher than initial, and after 30 days, the alpaca SSC decreased dramatically ( Results know showed ).…”

“…The DMEM medium has been used in the proliferation of SSCs from several species such as buffalo (Kala et al, ), goat (Bahadorani et al, ) or human (Sani et al, ). The STEMPRO medium is also used in the culture of SSC in human (Akhondi et al, ; Medrano et al, ) or mouse (Ishikura et al, ; Kanatsu‐Shinohara et al, ; Sato et al, ). Our results have demonstrated that alpaca spermatogonial stem cells proliferated in both media, DMEM and STEMPRO.…”

“…Although the comparison of different culture media for the proliferation of SSC has been studied in other species like human (Gat et al, ), this is the first work that analysed the proliferation of alpaca SSCs in different culture media. Medrano et al () observed that STEMPRO promotes proliferation of cryopreserved human SSC, while DMEM did not. Aponte et al, () used two culture mediums, STEMPRO and MEM, for bovine SSCs, and found a greater proliferation with the STEMPRO medium, without significant differences.…”

<p>Role of stem cells in fertility preservation: current insights</p>

“…54 Others also examined the efficiency of differential plating to select germ cells from TCSs, but did not find a difference in germ-cell numbers recovered from whole TCSs and differentially plated cells after 14 days of culture. 55 To improve SSC propagation, cell sorting prior to culture was further applied. Coculture of SSCs sorted by fluorescence-activated cell sorting based on their HLA -/EPCAM + phenotype onto inactivated somatic feeder cells resulted in putative SSCs coexpressing DDX4 and UTF1, although their proliferation rate was poor and no survival was found after 4 weeks.…”

“…Coculture of SSCs sorted by fluorescence-activated cell sorting based on their HLA -/EPCAM + phenotype onto inactivated somatic feeder cells resulted in putative SSCs coexpressing DDX4 and UTF1, although their proliferation rate was poor and no survival was found after 4 weeks. 55 Other phenotypic markers, ie, GFRα1, GPR125, SSEA-4, KIT -/ITGβ1 -, CD9, ITGα6, THY1, and FGFR3, have been used to select monkey or human SSCs, 38,43,44,[48][49][50][56][57][58][59][60][61][62][63][64][65] but among 16 studies, only 5 cultured the sorted SSCs. 38,[48][49][50]59 Lim et al succeeded in long-term culture of CD9-sorted spermatogonia onto laminin-coated plates, but reported a low proliferation rate (20,000-80,000 cells in 130 days).…”