A Novel Approach for the Derivation of Putative Primordial Germ Cells and Sertoli Cells from Human Embryonic Stem Cells

Abstract

Using human embryonic stem cells (hESCs), we describe a novel method for the rapid derivation and enrichment of cells that are comparable to primordial germ cells (PGCs) and Sertoli cells. The methodology described is based on modest changes to the growth conditions commonly used to expand hESCs and does not require genetic manipulation or complex three-dimensional culture. Remarkably, we have determined that simply reducing the size of cultured ESC colonies and manipulating the number of feeding cycles, results in the rapid emergence of cells that are comparable to migratory PGCs. Importantly, these cells can be monitored and purified on the basis of the expression of the chemokine receptor CXCR4. Under more stringent differentiating conditions these cells mature and upregulate the expression of specific germ cell markers. Importantly, this process is accompanied by the development of Sertoli-like support cells. Such cells normally provide trophic support and immunoprotection to developing germ cells and may have significant clinical utility in the prevention of graft rejection. The putative Sertoli-germ cell cocultures generated in this study may ultimately be developed to study and manipulate interactions and processes involved in human gametogenesis.

Amniotic fluid stem cell-based models to study the effects of gene mutations and toxicants on male germ cell formation

“…The efficiency of differentiation from ES or iPS cells into haploid male germ cells is generally reported to be low. [14][15][16][17][18][19][20][21][22][23][24] Although there is no evidence for the speculation that this efficiency might be higher using AFS cells, to our opinion, the here reviewed knowledge warrants testing this new stem cell type for their usability for the in vitro investigation of sperm production (Figure 1).…”

Use of pluripotent stem cells for reproductive medicine: are we there yet?

“…It was shown that increasing BMP4 concentration from 10 ng/ml to 100 ng/ml significantly improved VASA expression, especially upon addition of BMP4 together with BMP7 and BMP8b (Kee et al 2006). Other groups also demonstrated a culture-enrichment strategy by sorting for cells positive for early PGC markers such as cKIT, SSEA1 and CXCR4 (Tilgner et al 2008;Bucay et al 2009;Park et al 2009). Further differentiation of only this selected cell population led to increased germ cell gene expression (Tilgner et al 2008;Bucay et al 2009;Park et al 2009).…”

Productionin vitrode cellules germinales murines et humaines à partir de cellules souches pluripotentes

“…Certains marqueurs commencent à être reconnus comme incontournables pour l'établissement du phénotype des CGP : ainsi PRDM1, sous le contrôle de la signalisation BMP/SMAD, est essentiel à l'orientation des cellules CSE vers une identité CGP chez la souris comme chez l'homme. Chez l'humain, deux équipes ont mis en évidence les transcrits PRDM1 dans des CGP obtenues à partir de hCSE par différents protocoles qui seront détaillés ci-après [7,8] TRA-1-60 [7,16,22] TRA-1-81…”

“…[5, 7,13,16,19,22,24,25,27] Cellules Sertoli, cellules Leydig, cellules de la granulosa, cellules de la thèque…”

“…Follicle stimulating hormone receptor/ luteinizing hormone receptor [7,8,17] Cellules Sertoli, cellules de la granulosa…”