科研之友
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摘要
Cataract, the leading cause of blindness worldwide, is caused by the apoptosis of lens epithelial cells (LECs). alpha A-crystallin is a major structural protein of the lens. However, the antiapoptotic function of alpha A-crystallin in lens stem cells remains unclear. In this study, primary LECs were isolated from postnatal 3-5 days of SD rats and transfected by Sendai virus loaded with four factors, OCT3/4, Sox2, c-Myc, and Klf4, to induced pluripotent stem cells (iPSCs). LEC-iPSC-like cells were identified by immunofluorescent staining. Cry alpha A-specific shRNA lentivirus was used to knockdown alpha A-crystallin in LEC-derived iPSC-like cells, which were treated with tert-Butyl hydroperoxide. The apoptosis of LEC-iPSC-like cells was examined by flow cytometry. We reprogrammed LECs and obtained embryonic stem cell-like colonies. LEC-iPSC-like cells with normal karyotype expressed pluripotent markers such as SSEA-4, TRA-1-60, and TRA-1-81. Knockdown of alpha A-crystallin increased the apoptosis of LEC-iPSC-like cells and rendered them less resistant to oxidation stress induced by tert-Butyl hydroperoxide. In conclusion, LECs could be reprogrammed into iPSC-like cells and alpha A-crystallins could protect LEC-iPSC-like cells from oxidation stress-induced apoptosis.
- 出版日期2016-10
- 单位中南大学
