可注射性纤维蛋白胶联合肌源干细胞移植治疗女性压力性尿失禁的实验研究-外科学专业毕业论文.docxVIP

Experimental Experimental Study On Muscle—Derived Stem Cell Transplantation Combined With Injectable Fibrin Glue In Rat Models Of Female Stress Urinary Incontinence ABSTRACT Objeetive To explore the effect and mechanism of Muscle．Derived Stem Cells transplantation combined with injectable fibrin glue in rat models of f．emale stress urinary incontinence in order to provide a new cytotherapy method． Methods 1．The rat MDSCs were isolated and cultivated by modified preplating process．The expression of eell surface antigens was tested by flOW cytometry． immunofluorescence and immunohistochemical method respectively．The P3．MDSCs were cultivated by differentiation medium．The myotubes were identified by the expression of MvHC via immunofluorescence method． 2．The P3．MDSCs were seeded on sterile coverslips in 6．well plates with a density of 1×1 03／ml to culture inside three．dimensional condition and on surface of fibrin glue respectively．The morphology of MDSCs Was observated by scanning electron microscope at 1吼and 7m day of cultivation．The P3．MDSCs were seeded in 96．well plates with a density of 5×1 0’／well and then devided into three groups：A group (culture inside three．dimensional condition of fibrin glue)．B group(culture on surface of fibrin glue)，C group(culture on surface of 96-well plate)．The cell growth in different culture conditions was observed by a inverted microscope．The MTS method was used to test OD value of each group to assess the cell proliferation． 3．The P 1．MDSCS were infected by pGC．FU．GFP．Lv-a 1entiviral vector with green fluorescent protein(GFP)．at optimal MOI and then passaged．The GFP—positive cells were quantified by FACS．Pudendal nerve．transected(PNT、rats were utilized as stress urinary incontinence(SUI)models．Seventy five animals were divided into five groups(n=1 5)：Sham(S)，PNT+FG—injection(F)，vehic control(D)， PNT+MDSCs—injection(M)，and PNT+MDSCs plus FG—injection(FM)；And they were then split into 1、2 and 4 weeks subgroups．GFP．1abeling MDSCs wi