Leukemia Inhibitory Factor (LIF) Enhances Germ Cell Differentiation from Primate Embryonic Stem Cells

Abstract

Recently, several research groups have shown that germ cells can be produced in vitro from pluripotent embryonic stem cells (ESCs). In the mouse, live births of offspring using germ cells induced from ESCs in vitro have been reported. Furthermore, some efficient methods for inducing the useful number of germ cells from ESCs have also been developed. On the other hand, in primates, despite the appearances of germ cell-like cells including meiotic cells were observed by spontaneous differentiation or introducing transgenes, it has not been determined whether fully functional germ cells can be derived from ESCs. To elucidate the property for the germ cells induced from primate ESCs, specification of the promoting factors for the germ cell development and improving the efficiency of germ cell derivation are essential. Leukemia inhibitory factor (LIF) has been reported as one of the important factors for mouse primordial germ cell (PGC) survival in vitro. However, the effects of LIF on germ cell formation from pluripotent cells of primates have not been examined. The aim of this study is to determine whether LIF addition can improve in vitro germ cell production from cynomolgus monkey ESCs (cyESCs). After 8 days of differentiation, LIF added culture induced dome-shaped germ cell colonies as indicated by the intense expression of alkaline phosphatase activity (ALP). These cells also demonstrate high-level expression of the germ cell-marker VASA, OCT-4, and BLIMP-1, and show SSEA-1 expression that supports their early stage germ cell identity. Finally, we observed that adding LIF to differentiating cultures inhibited meiotic gene expressions and increased the percentage of ALP-positive cells, and demonstrate that the addition of LIF to differentiation media increases differentiation of early germ cells from the cyESCs.

LIF maintains progenitor phenotype of endothelial progenitor cells via Krüppel-like factor 4

“…LIF with VEGF maintained the preliminary, progenitor phenotype of EPCs LIF is a key cytokine in stem cell biology and combining with other growth factors, it upregulates embryonic germ (EG) colony number (Fukunaga et al, 2010;Tee et al, 2010). VEGF is the most important cytokine in EPCs and could increase EPC number and enhance proliferation in a dose-dependent manner (Fujinaga et al, 2009;Xiao-Yun et al, 2011;Zhou et al, 2008).…”

Germline development from human pluripotent stem cells toward disease modeling of infertility

“…The differentiation can be enhanced by conditioned medium from testicular or ovarian cells or by adding recombinant BMP4, RA or SCF (76). The self-renewal of differentiated PGC-like cells can be prolonged by LIF treatment (77). Further studies, including functional assays in vivo, would be interesting to examine their developmental potential and molecular mechanisms of primate germline development as a substitute model for human germ cell differentiation from pluripotent stem cells.…”

Direct Differentiation of Human Pluripotent Stem Cells into Advanced Spermatogenic Cells: In Search of an In Vitro System to Model Male Factor Infertility

“…First, human PSCs were differentiated into SSC-like cells, cells that reside at the foundation of spermatogenesis. Several previous studies have shown the ability of human and non-human primates PSCs to differentiate into PGCs [3,17,29,44,45,55,56,62]. Although this cell lineage has the capability of restoring fertility in rodents, including primordial germ cells (PGCs) derived from mouse PSCs [6,20], SSCs remain the gold standard for colonizing cells which recapitulate spermatogenesis following transplantation [1,27].…”