Distribution Of Mesenchymal Stem Cells Transfected With Hepatocyte Growth Factor In The Rat's Kidney With Unilateral Ureteral Obstruction

Obstructive nephropathy(ON) is a common disorder in urinary surgery,which major pathologic feature is renal interstitial fibrosis.In children,obstructive nephropathy is the primary cause for end stage renal disease(ESRD) and represents 16.1%of all pediatric kidney transplantation.Bone marrow-derived mesenchymal stem cells(MSCs) are characterized with self proliferation and multi-lineage differentiation.MSCs can differentiate to histocyte originated from mesoderm,endoderm and neuroectoderm,include osteoblast, cardiocyte,nerve cell and renal cell.It has been shown that systemic delivery of MSCs could protect mice with acute renal failure from renal function impairment and severe tubular injury.Hepatocyte growth factor(HGF) as a determiner facilitate adult stem cells differentiate to renal cell,and posses the ability to prevent apoptosis of renal tubular cell.In this sdudy,bone marrow-derived MSCs of SD rats were separated and cultured in vitro,and then transfected with Ad-HGF.Rats model with unilateral ureteral obstruction(UUO) were established and Ad-HGF transfected MSCs were injected into the rats with UUO.Finally,we observe whether MSCs transfected with HGF can immigrate and localize in the rat's kidney with UUO,which may provide a theoretical clue for gene and cell therapy for obstructive nephropathy.Materials and methodsBone marrow derived-MSCs of the male SD rats were separated and cultured in vitro,and the phenotypic identification of CD34 and CD90 were detected by flow cytometry.MSCs were transfected with Ad-GFP in different MOI in order to select an optimal MOI.Transfection efficiency was detected by flow cytometry.The expression of HGF in MSCs transfected with optimal MOI was measured with ELISA assay.Sixteen female SD rats with UUO were randomly divided into two groups: transplantation group and control group.Ad-HGF transfected MSCs or saline were injected into the rats by vena caudalis.Kidney tissue of the rats were collected at the end of the 7th day and 14th day after operation.The distribution of Y chromosome in the kidney was determined by in situ hybridization.Results1.After 7-9 days,primary MSCs can be passaged.After generation,cell proliferation rate increase,after 4-5 days,MSCs can be passaged again.MSCs surface antigen by flow cytometry show CD34(—),CD90 99%.2.Transfection efficiency of Ad-GFP to MSCs is 73.29%when MOI equaled 400.3.The expression of HGF in MSCs reached the highest level after being transfected with Ad-HGF for 48h.4.Y chromosome positive cells were found only in the obstructed kidney of the transplantation group.The positive cells mainly distributed in the tubular cells.Conclusions1.We can isolate and purify MSCs by means of attachment culture method.2.MSCs were ideal genetic carrier for HGF gene therapy.3.MSCs transfected with HGF can immigrate to the rat's kidney with UUO,and mainly distributed in the region of renal tubular epithelial cells.