The Repair Of Bladder Defects In Rabbits With Adipose Derived Stem Cells Combined With Xenogeneic Bladder Acellular Matrix

Objective:(1)Studying the feasibility of CM-Dil markers of adipose derived stem cells,and observe the fluorescence labeled adipose stem cells and bladder acellular matrix forming cell co culture of scaffold complex expression,to ensure the quality of transplanted cells and the effect of tracer labeled cells.(2)Evaluate the cell scaffold composite for repair of rabbit bladder defect model,observation and identification of cell scaffold composite repair at the expression of specific markers.Methods: In the bilateral inguinal adipose tissue of New Zealand white rabbit,the primary Adipose derived stem cells were obtained after treatment,osteogenic and adipogenic induction staining and flow cytometry detection of CD29,CD34,CD44 and CD45 were used to identify the adipose derived stem cells,and preparation of rat BAM.Adipose derived stem cells(P3)were labeled with CM-Dil,and the adipose derived stem cells were seeded on the BAM surface by two precipitation method.The fluorescence expression of cells and cell scaffold complexes was observed and evaluated.Surgical implant cell scaffold to observe the growth of rabbit bladder defect model;respectively after 2,4,6,8 weeks to obtain experimental animal bladder repair,selected location,were made into frozen and paraffin sections to observe the fluorescence expression of frozen tissue sections using fluorescence microscopy,expression by immunohistochemical method detection of specific structure components of smooth muscle cell marker.Results:(1)The third generation of rADSCs fluorescence labeled CM-Dil stable expression,there are still strong GFP expression by cell scaffold co cultured with rat BAM.(2)Two rabbits died after 7d and 16 d,because of leakage of urine;expression of tissue fluorescence weakened but still,with no well-defined repair position,Masson staining found that organizational structure is complete,the muscle fiber was significantly,immunoh istochemical expression of smooth muscle cell specific markers: a-SMA.Conclusion:(1)The use of CM-Dil as a method of fluorescent labeling of cells is feasible,the expression level of in vitro and in vivo fluorescence were satisfactory;(2)The bladder acellular matrix(BAM)in experimental rabbits after 8 weeks can be abs orbed and degraded,is a good scaffold materials in vivo,rADSCs can differentiate into smooth muscle cells.rADSCs and BAM can be used to construct tissue-engineered bladder in vitro by static inoculation,which is an ideal bladder repair and reconstruction material for repairing the defect of rabbit bladder.