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The transcriptome data of pig embryonic stem cells cultured without feeder cells

ABSTRACT: The transcriptome data of pig embryonic stem cells cultured without feeder cells

PROVIDER: PRJNA787086 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17167227_1.fastq.gz	Fastqsanger.gz
	SRR17167227_2.fastq.gz	Fastqsanger.gz
	SRR17167228_1.fastq.gz	Fastqsanger.gz
	SRR17167228_2.fastq.gz	Fastqsanger.gz
	SRR17167229_1.fastq.gz	Fastqsanger.gz

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The transcriptome data of pig embryonic stem cells cultured without feeder cells

Project description:Here, we examined the effect of LDN-193189, an inhibitor of the BMP signaling pathway, on pig embryonic stem cells (ESCs) in the absence of feeder cells to develop a feeder-free culture system. Feeder cells play an important role in supporting the pluripotency of pig ESCs by blocking trophoblast and mesodermal differentiation through inhibition of the BMP pathway. Transcriptome analysis revealed that LDN-193189 treatment instead of coculturing with feeder cells could help maintain the pluripotency and homogenous cell population of pig ESCs by stimulating the secretion of chemokines and suppressing differentiation. We confirmed that LDN-193189-treated pig ESCs cultured without feeder cells maintained pluripotency for an extended period and were capable of differentiating in vivo and in vitro. The pig ESCs represented distinct characteristics of pig epiblasts as an in vivo counterpart at a transcriptomic level. In summary, we developed feeder-free culture systems for pig ESCs, and our findings will improve the understanding of pluripotency gene networks and comparative embryogenesis.

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Project description:The pluripotency maintenance of pluripotent stem cells (PSCs) cultured in vitro requires the suitable microenvironment, which is commonly provided by the feeder layer. However, the preparation of feeder layer is time consuming and labor exhaustive. More importantly, the feeder cells treated with mitomycin or gamma-ray irradiation brings heterologous contamination to stem cells. The feeder-free PSC cultures are associated with high costs because of the requirement for additional supplements and special media. In this study, we characterize the pluripotency and metabolic status of bovine ESCs-F7 (classic bESCs lines, abbreviated as F7), which were cultured on methanol fixed mouse embryonic fibroblasts (MT-MEFs) or mitomycin C treated MEFs (1M-MEFs). MT-MEFs could be reused several times and were highly resistant to digestive enzymes. The relative expression levels of pluripotent markers were different between F7 cultured on MT-MEFs (marked as MT-F7) and those cultured on the 1M-MEFs (1M-F7). The long-term cultured MT-F7 cells formed embryoid bodies in vitro, showing the ability to differentiate into endodermal, ectodermal, and mesodermal germ layers like 1M-F7. RNA-sequencing analysis showed that the replacement of the feeder layer from 1M-MEFs to MT-MEFs lead to a novel steady transition of the F7, which included alteration of the expression patterns of genes that regulate pluripotency and metabolism. Further, the long-term cultured bovine expanded pluripotent stem cells (bEPSCs) on MT-MEFs (MT-bEPSCs) formed classical colonies, maintained pluripotency, and demonstrated elevated level of metabolic activity. In conclusion, this study demonstrated that methanol-fixed MEFs were efficient feeder layer that maintain the unique pluripotency and the distinctive metabolic characteristics of the bPSCs cultured in vitro.

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