Microarray analysis of embryo-derived bovine pluripotent cells: The vulnerable state of bovine embryonic stem cells

Abstract

Although there are many studies about pluripotent stem cells, little is known about pluripotent pathways and the difficulties of maintaining the pluripotency of bovine cells in vitro. Here, we investigated differently expressed genes (DEG) in bovine embryo-derived stem-like cells (eSLCs) from various origins to validate their distinct characteristics of pluripotency and differentiation. We identified core pluripotency markers and additional markers which were not determined as pluripotency markers yet in bovine eSLCs. Using the KEGG database, TGFβ, WNT, and LIF signaling were related to the maintenance of pluripotency. In contrast, some DEGs related to the LIF pathway were down-regulated, suggesting that reactivation of the pathway may be required for the establishment of true bovine embryonic stem cells (ESCs). Interestingly, oncogenes were co-down-regulated, while tumor suppressor genes were co-up-regulated in eSLCs, implying that this pattern may induce abnormal teratomas. These data analyses of signaling pathways provide essential information on authentic ESCs in addition to providing evidence for pluripotency in bovine eSLCs.

Efficient derivation of stable primed pluripotent embryonic stem cells from bovine blastocysts

“…Most, if not all, of the reported bESC lines do not pass standard pluripotency tests, i.e., in vitro embryoid body formation, in vivo teratoma assay, and/or chimera formation. Moreover, they show poor derivation efficiencies, limited proliferation capacities, and loss of pluripotency markers after extensive passages (6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16).…”

Bovine Pluripotent Stem Cells: Current Status and Prospects

“…Unlike in mice, signaling between LIF and STAT3 may be disconnected in cattle, which is possibly the reason why the true bESCs could not be generated in the previous culture system using LIF. This suggests that STAT3 may be the key molecule in establishing bESCs [24]. Inactivation of the WNT pathway is essential for the development of preimplantation and early post-implantation embryos and stabilizes the cytoplasmic β-linker proteins by inhibiting GSK-3β.…”

Strategy to Establish Embryo-Derived Pluripotent Stem Cells in Cattle

“…To understand pluripotency, several transcriptome studies have also been conducted in cultured cells from bovine blastocysts. With various blastocyst-derived cells, microarray results suggested signaling pathways for bovine pluripotency [ 44 ]. Three different putative bESCs from IVP, NT, and PA embryos were used, with the findings revealing common increases in TGF-β, Wnt, and LIF signaling pathways related to pluripotency, implying that these pathways may be pivotal for capturing authentic pluripotency in bovines.…”

“…There are mutations and cell instability that occur in the artificial process of generating and culturing stem cells. In addition, the established stem cells still have the epigenetic memory and methylation status of the donor cells, which influence cell differentiation and fate determination [ 44 , 107 , 108 ]. Although gene editing, including CRISPR technology and epigenetic modifiers, are generally utilized, many unknown issues still remain, including side effects such as off-targeting.…”