The Function Of Bone Marrow Stromal Stem Cells After Tansdifferentiated Into Renal Tubular Epithelial Cells In Vitro And The Intervention Of Huangqi

Objective To investigate whether bone marrow stromal stem cells (BMSCs) have part functions of renal tubular epithelial cells (RTECs) after transdifferentiated into RTECs in vitro,providing an experimental basis for the clinical application of BMSCs.Observing the effects of the Chinese herb of HuangQi in the process of BMSCs transdifferentiate into RTECs. Methods①Isolate, culture and identification BMSCs and RTECs of BALB/C mice.②To establish a mouse renal acute ischemia-reperfusion injury (AIRI) model by occlusion bilateral renal artery.③BMSCs were co-cultured with RTECs of normal mice and AIRI mice in vitro, and add to intervene with the traditional Chinese medicine Huang Qi.④After the induction of co-culture, BMSCs identified by immunohistochemistry, the Expression of angiotensinⅡreceptor (AngⅡ) and Endothelin-1 (ET-1)) of BMSCs were detected by quantitative PCR and biochemical measure the induced BMSCs on the urea nitrogen and uric acid transporter levels to know whether there had been transdifferentiation of BMSCs into RTECs and has part function of RTECs.Results①After 24 hours the inoculation BMSCs could adhere to the flask,where single-dispersal cell and sheet-shaped cell clone were formed.The third generation cells were purified and most of them were spindle-shaped.They showed vortex shaped or parallel arrangement growth. Identification of BMSCs by flow cytometry shows CD29 expression was strongly positive, CD45, CD34, ClassⅡnegative expression. (2)The level of AIRI mice creatinine, urea nitrogen begins to rise, and reached the peak at 24h, renal pathology showed the performance of acute tubular necrosis.③After induction, BMSCs express specific marker proteins of RTECs CK18, whereas the expression of Vimentin and a-SMA decreased,the gene expression of AngⅡreceptor and ET-1 of BMSCs significantly increased detected by quantitative PCR, HuangQi induced group also increased.The function of BMSCs to transfer BUN was no significant change, While the function of transporting UA significantly increased, Huang Qi induced group the most obvious. Conclusion Non-invasive vascular clip bilateral renal artery occlusion in mice can successfully establish a mouse model of kidney AIRI; BMSCs can be isolated,cultured and amplificated in vitro;RTECs co-cultured with BMSCs in vitro can induce BMSCs transdifferentiating into RTECs; has part function of RTECs; HuangQi has facilitating role in transdifferentiation of BMSCs in vitro.