Abstract:
Objective We aimed to improve the stemness of chicken induced pluripotent stem cells using the Pcgf2 gene together with conventional reprogramming factors to induce chicken pluripotent stem cells.
Method Six conventional reprogramming factors including Oct4, Sox2, Nanog, Lin28, C-myc and Klf4 as well as Pcgf2 gene were amplified and reconstituted with lentiviral vectors as lentiviral plasmids using chicken early embryos as templates. After packaging the lentivirus and measuring the corresponding viral titers, chicken embryonic fibroblasts were infected in groups and stem cell markers were assayed during the induction process.
Result The cells induced by the combination of conventional reprogramming factors and Pcgf2 all expressed phosphatase activity early. Inducing the pluripotent stem cells with Pcgf2 could make the cells appear stem-like morphology earlier, and the expressions of endogenous genes Oct4, Sox2, Nanog and Lin28 increased more stably.
Conclusion Pcgf2 can effectively enhance the reprogramming of cells to approach the stem cell state after being added to the induction system. The research provides a theoretical reference for the selection of reprogramming factors to induce complete reprogramming of cells in chickens and the mechanism study.
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