Optimized effect of connexin 43 gene silencing on the proliferation of fetal liver stem cells

doi:10.3969/j.issn.2095-4344.2015.14.001

Abstract

Abstract:

BACKGROUND: Fetal liver stem cells have the potential to differentiate into hepatocytes and bile duct cells, and participate in the repair and reconstruction of the liver, which is an important source of hepatocytes. But there are a little amount of fetal liver stem cells in human body, and how to obtain a certain number of high-purity fetal liver stem cells is currently a hot research.

OBJECTIVE: To construct a siRNA carrier that can effectively inhibit the expression of connexin 43 (Cx43) in rat fetal liver stem cells, and to investigate the effect of Cx43 inhibition on the proliferation and cell cycle of fetal liver stem cells cultured in vitro.

METHODS: Fetal liver stem cells were cultured by the suspension culture in vitro, siRNA sequences targeting Cx43 (Cx43-siRNA) and negative control sequence (NC-siRNA) were designed and synthesized. Then, rat fetal liver stem cells were transferred electrophoretically and divided into three groups: blank group, NC-siRNA group, Cx43-siRNA group. Real-time PCR and western blot were used to assess the knockdown efficiency. Cellular proliferation was determined by cell growth curve and cell counting kit-8 assay. The cell cycle was analyzed by flow cytometry.

RESULTS AND CONCLUSION: After transfection, the Cx43 gene and protein expression levels were declined dramatically in the Cx43-siRNA, NC-siRNA and blank groups, and the cells grew faster. The number of cells at G₀/G₁ phase decrease, but the number of cells in S phase increased. There were significant differences between the groups (P < 0.05). Electrophoretic transfer of Cx43-siRNA can promote the proliferation of cultured fetal liver stem cells and optimize the cell culture.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Hepatocytes, Connexin 43, RNA, Small Interfering, Cell Proliferation

CLC Number:

R394.2

Zhang Zeng-guang, Qin Ming-fang. Optimized effect of connexin 43 gene silencing on the proliferation of fetal liver stem cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(14): 2133-2137.

Figures/Tables 4

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