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Wnt signaling associated small molecules improve the viability of pPSCs in a PI3K/Akt pathway dependent way
Author(s) -
Li Yan,
Wu Shuang,
Li Xuechun,
Guo Shimeng,
Cai Zhuang,
Yin Zhi,
Zhang Yu,
Liu Zhonghua
Publication year - 2020
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.29514
Subject(s) - wnt signaling pathway , embryonic stem cell , microbiology and biotechnology , induced pluripotent stem cell , pi3k/akt/mtor pathway , biology , viability assay , stem cell , cell , protein kinase b , regenerative medicine , cellular differentiation , chemistry , signal transduction , biochemistry , gene
Although we have obtained porcine pluripotent stem cell lines (pPSCs) from blastocysts, the cells exhibit flat clonal morphology and do not support single‐cell passage. There is massive cell death after cell dissociation, and the efficiency of single‐cell colony is generally ≤10%. In a recent study, we got a new pPSCs using two Wnt signaling pathway regulators CHIR99021 and XAV939. This cell had strong biological viability, small‐domed morphology, and its cloning efficiency after dissociation was 80–90%. The CH/XAV‐treated cells expressed elevated levels of pluripotent genes, and possessed differentiation abilities both in vitro and in vivo, proven by the formation of embryonic bodies and teratomas with three germ layers. Furthermore, we found that the combinative use of CHIR99021 and XAV939 resulted in β‐catenin‐maintained expression in the cytoplasm but not translocation to the nuclei for WNT/TCF activation. In the meanwhile, E‐cadherin located on the cell membrane, thereby activated the PI3K/Akt signaling pathway to enhance the pluripotency of the cells. Our study obtained new pPSCs, which were even closer to the naïve state with only two small molecule inhibitors, and the improved pluripotency of pPSCs could facilitate transgenic manipulation and regenerative medicine research. Besides, our study casted a light on the understanding of pPSCs and the derivation of authentic porcine embryonic stem cells.