Effect Of Bone Mesenchymal Stem Cells Repair Immature Rat Chronic Renal Insufficiency

Objective:To study the effect of allogenic transplatation of Bone Mesenchymal Stem Cells(BMSCs) repair immature rat Chronic Renal Insufficiency(CRI).Methods:1.The culturing,identification and label of BMSCs in vitro: First got male Wistar rat weighting 40~60g and killed it off neck after narcotized,soaked in 75% alcohol for 15 minutes,put in a aseptic plate,then washed out the whole bone warrow by L-DMEM,cultured these cells by the whole bone marrow culture method combined with centrifugation,then picked up the third generation cells which were at good growth status and detected the expression of CD45 and CD90 through FCM.Third using Adipogenic and Osteogenic Differentiation Medium induce the cells differentiate into adipocytes and osteoblasts to identify the multiple potential differentiation by the oil Red O and Alizarin red staining.2.Grouping and model establishment of rats:A total of 90 male Wistar rats weighting(91±12)g were randomly assigned to three groups :BMSCs-treated group(group A):35 rats:CRI group(group B):35 rats and Normal control group(group C):20 rats.Rats in group C were intragastric administration with distilled water while group A and B were given adenine suspension 100mg/(kg?d) for 5 weeks to establish the model of CRI.3.The effect of BMSCs in CRI :At 24 hours following the CRI model were established,the A group received an injection of BMSCs(1×106cells)by DAPI labeled via tail vein, while group B and group C received PBS(1ml). On the days of 3,7,14,28 and 42 after BMSCs were injected,the BUN,serum creatinine,24-h urine protein were measuredand renal morphologic changes were observed through HE and PAS staining.Results:1.Culturing,identification and label of BMSCs in vitro: The third generation BMSCs were arranged orderly and had the typical spindle-shape. CD90 expressed and CD45 was not by flow-cytometry frequently.Red lipid droplets and calcified nodules formed after the Adipogenic and Osteogenic Differentiation Medium induced.2.Grouping and model establishment of rats:Compared with C group, group A and B after adenine suspension 100mg/(kg?d) for 5 weeks,The outcomes of renal function: There were no significant differences between the group A and the group B in the blood urea nitrogen,serum creatinine,24 h urine protein and renal morphologic changes before BMSC transplantation at 28 days;At 42 days these indicators of group A were decreased significantly than group B.3.Renal histology:There were no significant differences between the group A and the group B before 28 days. All the cases demonstrated glomerular hypertrophy,renal capsule expansion,basement membrane thickening,mesangial expansion,and the outline of renal tubules was not clear, renal tubules arranged unorderly,the epithelial cells of renal tubular were swollen or cleaved,even necrosis,cell matrix increased,the protein tube deposition appeared in renal tubular,the inflammatory cells infiltrated significantly,but no evident local glomerular sclerosis and tuberous sclerosis presented. The inflammatory cells infiltrated significantly in interstitium.At 42 days,the outline of renal tubular were clearer in group A than group B,the epithelial cells swelling reduced,and inflammatory cells decreased significantly.Conclusions:1.The whole bone marrow culture method combined with centrifugation could culture BMSCs at good status.2.Mouse-anti-rat CD45 and CD90 can identify bone marrow mesenchymal stem cellssimple and practically,and the cells were insured as BMSCs through the multipotent capability differentiation of adipogenic nutrition liquid osteogenic and nutrition liquid.3.That adenine suspension 100mg/(kg?d)fed rats for 5weeks could establish the model of CRI successfully;4.BMSCs could improve the CRI of rats significantly,repair renal tubular structure and inhibite the infiltration of inflammatory cells.