[关键词]
[摘要]
目的: 探索大鼠膀胱无细胞基质负载骨髓间充质干细胞体外构建组织工程膀胱复合物的可行性。
方法: 全骨髓贴壁培养法提取大鼠骨髓间充质干细胞( BMSCs) ，物理和酶消化法相结合制备大鼠膀胱无细胞基
质( BAMG) ，将分离纯化的P3 代BMSCs 接种在BAMG 上共同培养构建组织工程膀胱复合物，HE 染色观察复合
物的细胞生长情况，以P3 代BMSCs 的单独培养做对照，绘制两组细胞生长曲线图，对比评价BAMG 对BMSCs
生长有无影响。结果: ( 1) 全骨髓贴壁培养法分离培养的BMSCs 经流式细胞仪检测细胞表面标志物CD29、CD44
抗原表达阳性，而CD34 呈阴性表达。( 2) 应用物理法和酶消化法相结合成功制备BAMG，HE 和扫描电镜均未
见细胞残留，具有完整的胶原及纤维结构。( 3) BMSCs 与BAMG 能黏附生长，体外成功构建组织工程膀胱复合
物，其生长曲线和对照组基本相同。结论: 全骨髓贴壁培养法能简单有效的增殖纯化BMSCs，物理法和酶消化法
相结合可以成功制备BAMG，二者体外成功构建组织工程膀胱复合物。
[Key word]
[Abstract]
Objective: To explore the construction feasibility of mouse tissue engineering bladder
complex with bone marrow mesenchymal stem cells( BMSCs) seeded on bladder acellular matrix graft
( BAMG) ．Methods: The mouse BMSCs were obtained with the method of whole bone marrow adherent，
BAMG was produced through the combined methods of physical digestion and enzymatic digestion．P3
generation BMSCs were seeded on BAMG and co－cultured．The growing situation of BMSCs were tested
by the staining method of HE． As the control group，P3 generation BMSCs were cultured alone． The
growth curves between two groups were drawn and to evaluate if BAMG had any effect of BMSCs．Results:
( 1) BMSCs’surface markers expressed CD29，CD44 antigen positive，a negative and CD34 expression
detected by flow cytometry instrument; ( 2) BAMGs were not seen cells residual by HE and
scanning electron microscopy ( SEM) and had whole collage and fibrous structure; ( 3) BMSCs and
BAMG can stick to grow， in vitro to construct tissue engineering bladder complex，and had the same
effect with the control group．Conclusion: The whole bone marrow adherent culture was a simple and effective
method of purification and proliferation of BMSCs，and the combination of the physical and enzymatic
digestion could be successfully used to produce BAMG，BMSCs could be seeded on BAMG， they
can construct tissue engineering bladder complex together．
[中图分类号]
R697．3
[基金项目]
收稿日期: 2016－01－10; 修回日期: 2016－04－02
基金项目: 内蒙古自治区自然科学基金资助项目( 2011BS1115)
作者简介: 高红亮( 1982－) ，男，内蒙古医科大学2013 级在读硕士研究生。
通讯作者: 易发现，主任医师，E－mail: faxian．yi@ yahoo．com 内蒙古医科大学附属医院泌尿外科， 010050
