Chemically Defined Media Can Maintain Pig Pluripotency Network In Vitro

Abstract

Summary Pig embryonic stem cells (pESCs) have been considered an important candidate for preclinical research on human therapies. However, the lack of understanding of pig pluripotent networks has hampered establishment of authentic pESCs. Here, we report that FGF2, ACTVIN, and WNT signaling are essential to sustain pig pluripotency in vitro . Newly derived pESCs were stably maintained over an extended period, and capable of forming teratomas that contained three germ layers. Transcriptome analysis showed that pESCs were developmentally similar to late epiblasts of preimplantation embryos and in terms of biological functions resembled human rather than mouse pluripotent stem cells. However, the pESCs had distinct features such as coexpression of SSEA1 and SSEA4, two active X chromosomes, and a unique transcriptional pattern. Our findings will facilitate both the development of large animal models for human stem cell therapy and the generation of pluripotent stem cells from other domestic animals for agricultural use.

A cytokine screen using CRISPR-Cas9 knock-in reporter pig iPS cells reveals that Activin A regulates NANOG

“…But, patterns of NANOG in pig pluripotent stem cells (PSCs) from pre-implantation embryos are different from pig iPS cells. NANOG is highly expressed in pig ES cells from ICM/epiblast [13] and in pig EPS cells [12]. The NANOG expression of pig iPS cells in doxycycline-inducible system with 4 factors (OSKC) was low [36].…”

“…NANOG is also highly expressed in pig preimplantation embryos [10]. Recently, pig pluripotent stem cells (PSCs) were established from the inner cell mass of pig blastulas [11][12][13]. We found that induced pluripotent stem cells (iPSCs) from pigs express NANOG heterogeneously [14], as in mouse PSCs [15,16].…”

“…Pig pluripotent stem cells can be used in pig breeding, to model pig disease and to test pre-clinical regenerative medicines. Although pig expanded pluripotent stem cells [12], dome-shaped iPS cells [14], and ESCs [13] have recently been established, germline ESCs/iPSCs are not yet available. Many cytokines have been used to generate pig pluripotent stem cells, such as LIF [32][33][34], bFGF [35], and LIF and bFGF in combination [36,37].…”

Pig Pluripotent Stem Cells as a Candidate for Biomedical Application

“…Several studies showed that ESCs were more efficient for cloning of mouse than differentiated cells such as immune cells, neuron, and fibroblasts (Hochedlinger and Jaenisch, 2002). which indicates that these cells could have the potential for preclinical study of human cell therapy (Choi et al, 2019). And they found lipid supplementation was crucial for maintaining pig pluripotency in vitro, which indicated that metabolic niche also affects deriving and maintaining pluripotency in the ungulate.…”

“…To accomplish these goals with pig PSCs, there are hurdles remain to be overcame. First of all, although recent study represented authentic pig ESCs can derive using chemically defined media (Choi et al, 2019), it is required to develop more defined culture conditions such as feeder-free culture and reprogramming method for pig PSCs. Analysis of enhancer showed that different transcription factors were involved in the expression of OCT4 known as a core gene of pluripotency networks , which means a majority of pig pluripotency circuit remains to be elucidated.…”