All-Trans Retinoic Acid Combined With Type Iv Collagen Promotes Differentiation Of Embryonic Stem Cells Into Smooth Muscle Cells
Keywords
Embryonic Stem Cells All-Trans Retinoic Acid Type Iv Collagen Sm-¦¡ Actin Sm-Mhc
Abstract
Objective To explore whether all-trans retinoic acid (atRA) combined with type IV collagen (collagen IV) can better promote the differentiation of mouse embryonic stem cells (mESC) into smooth muscle cells. Methods mESCs were cultured in vitro, and the totipotency of mESCs was detected through alkaline phosphatase staining, SSEA1 staining and in vivo tumorigenesis experiments. Embryonic bodies (EBs) were prepared and suspended for 3 days and divided into: no induction (control group), atRA induction group, type IV collagen induction group, and atRA combined with type IV collagen induction group to allow them to adhere and differentiate. The differentiation of cells in different groups into smooth muscle cells was observed and compared through Western blot detection of smooth muscle cell antibody (SM-¦Á actin) and immunofluorescence double staining results of SM-¦Á actin and smooth muscle myosin heavy chain (SM-MHC). Compare. The results showed that the culture of mouse embryonic stem cells was successful after mESC totipotency identification. It was found that the SM-¦Á actin protein expression of cells in the atRA combined with type IV collagen induction group was significantly higher than that of the other three groups on the 7th and 14th days. At the same time, the SM-¦Á actin and SM-MHC double fluorescence staining on the 14th day was observed. It was found that the expression of SM-¦Á actin and SM-MHC in the atRA combined with type IV collagen induction group was also the highest, indicating that the differentiation and maturity of the cells was the best, followed by the atRA induction group and type IV collagen induction group, and the control group was the worst. Conclusion All-trans retinoic acid combined with type IV collagen can better promote the differentiation of embryonic stem cells into smooth muscle cells.
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Original research was done by Liu Danan, Zhang Hongming, Peng Chengfei, Tian Xiaoxiang, Shao Xiaoping, Yan Chenghui, Han Yaling
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