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猪胚外胚内细胞的特征分析。

Characterization of porcine extraembryonic endoderm cells.

机构信息

College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering and Technology, Northwest A&F University, Yangling, China.

Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.

出版信息

Cell Prolif. 2019 May;52(3):e12591. doi: 10.1111/cpr.12591. Epub 2019 Mar 21.

DOI:10.1111/cpr.12591
PMID:30896067
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6536407/
Abstract

OBJECTIVES

To date, many efforts have been made to establish porcine embryonic stem (pES) cells without success. Extraembryonic endoderm (XEN) cells can self-renew and differentiate into the visceral endoderm and parietal endoderm. XEN cells are derived from the primitive endoderm of the inner cell mass of blastocysts and may be an intermediate state in cell reprogramming.

MATERIALS AND METHODS

Porcine XEN cells (pXENCs) were generated from porcine pluripotent stem cells (pPSCs) and were characterized by RNA sequencing and immunofluorescence analyses. The developmental potential of pXENCs was investigated in chimeric mouse embryos.

RESULTS

Porcine XEN cells derived from porcine pPSCs were successfully expanded in N2B27 medium supplemented with bFGF for least 30 passages. RNA sequencing and immunofluorescence analyses showed that pXENCs expressed the murine and canine XEN markers Gata6, Gata4, Sox17 and Pdgfra but not the pluripotent markers Oct4, Sox2 and TE marker Cdx2. Moreover, these cells contributed to the XEN when injected into four-cell stage mouse embryos. Supplementation with Chir99021 and SB431542 promoted the pluripotency of the pXENCs.

CONCLUSIONS

We successfully derived pXENCs and showed that supplementation with Chir99021 and SB431542 confer them with pluripotency. Our results provide a new resource for investigating the reprogramming mechanism of porcine-induced pluripotent stem cells.

摘要

目的

迄今为止，人们已经做出了许多努力来建立猪胚胎干细胞（pES），但都没有成功。胚外内胚层（XEN）细胞可以自我更新并分化为内脏内胚层和滋养外胚层。XEN 细胞来源于囊胚内细胞团的原始内胚层，可能是细胞重编程的中间状态。

材料和方法

从猪多能干细胞（pPSCs）中生成猪 XEN 细胞（pXENCs），并通过 RNA 测序和免疫荧光分析进行鉴定。研究了 pXENCs 在嵌合小鼠胚胎中的发育潜能。

结果

成功地在补充 bFGF 的 N2B27 培养基中扩增了来自猪 pPSCs 的猪 XEN 细胞，至少传代 30 代。RNA 测序和免疫荧光分析表明，pXENCs 表达了鼠和犬 XEN 标志物 Gata6、Gata4、Sox17 和 Pdgfra，但不表达多能标志物 Oct4、Sox2 和 TE 标志物 Cdx2。此外，这些细胞在注入四细胞期小鼠胚胎时有助于形成 XEN。补充 Chir99021 和 SB431542 可促进 pXENCs 的多能性。

结论

我们成功地获得了 pXENCs，并表明补充 Chir99021 和 SB431542 可赋予它们多能性。我们的结果为研究猪诱导多能干细胞的重编程机制提供了新的资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/2744780a8fb2/CPR-52-e12591-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/655e59e077e0/CPR-52-e12591-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/0a1fce9e6a39/CPR-52-e12591-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/a83974ac07c6/CPR-52-e12591-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/2744780a8fb2/CPR-52-e12591-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/655e59e077e0/CPR-52-e12591-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/0a1fce9e6a39/CPR-52-e12591-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/a83974ac07c6/CPR-52-e12591-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/091e/6536407/2744780a8fb2/CPR-52-e12591-g004.jpg

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