胎儿肠壁结缔组织诱导骨髓间充质干细胞向肠上皮细胞的分化  

Bone marrow-derived mesenchymal stem cells differentiate into epithelial cells induced by fetal intestinal connective tissue

作　　者：段江洁[1] 王蓓[1] 张建华[2] 姜蓉[1] 汪维伟[1] 

机构地区：[1]重庆医科大学基础医学院干细胞与组织工程研究室,组织胚胎学教研室,重庆400016 [2]重庆医科大学临床学院妇产科,重庆400016

出　　处：《第三军医大学学报》2008年第18期1718-1721,共4页Journal of Third Military Medical University

基　　金：重庆医科大学创新基金(CX200307)~~

摘　　要：目的观察胎儿肠壁结缔组织能否诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分化为上皮细胞。方法制备含胎儿肠黏膜和黏膜下层的组织块,酶消化去肠上皮。将DAPI标记的第3代BMSCs种植于组织块黏膜下层面,设加和不加EGF的A、B组;另设BMSCs细胞爬片的C、D对照组。体外培养至第12天,做光镜、CLSM的形态、组化和免疫组化观察。结果①荧光显微镜下组织块表面有DAPI荧光标记细胞,同一切片HE染色显示这些细胞呈上皮样形态。②免疫组化染色显示A、B组组织块表面的细胞和C组的细胞都能表达CK及CK20,D组为阴性;肠组织块有EGF表达。CLSM下组织块表面可见蓝色DAPI和红色CK或者绿色EMA双荧光标记阳性细胞。③PAS反应显示在细胞与结缔组织之间有基膜样结构出现。结论肠壁结缔组织能诱导BMSCs分化为上皮细胞,内源性EGF可能是其诱导因素之一。Objective To observe the possibility that bone marrow mesenchymal stem cells （BMSCs） induced by fetal intestinal connective tissue differentiate into epithelial cells. Methods The fetal intestinal tissue lumps composed of mucosa and submucosa were prepared with informed consent and approval of the Ethics Committee of our university, and the intestinal epithelium was removed with enzymatic digestion. The 3^rd generation cells of fetal BMSCs labeled with DAPI were planted on the surface of the bowel submucosa, and divided into group A （EGF added） and group B （without EGF addition）. BMSCs attached to cover glass were set as control groups C and D. After BMSCs were cultured in vitro for 12 d, their morphological, histochemical and immunohistochemical analysis was performed under light microscope and CLSM. Results The DAPI labeled cells could be seen on the surface of tissue lumps by fluorescence microscope, and in the same section stained by HE, these cells looked morphologically like epithelial cells. Immunohistochemical staining showed that the cells on the surface of tissue lumps in groups A, B and C expressed CK and CK20, but not in group D. EGF expressed in intestinal connective tissues. The double fluorescent labelling cells with blue DAPI and red CK or green EMA could be seen on the surface of tissue lumps observed by CISM. The basal lamina-like structure appeared between the cells and connective tissues in PAS reaction. Conclusion The intestinal connective tissues could induce BMSCs to differentiate into epithelial cells, and the endogenous EGF may be one of factors in this process.

关 键 词：肠 结缔组织 诱导 骨髓间充质干细胞 分化 

分 类 号：R321.5[医药卫生—人体解剖和组织胚胎学] R322.45[医药卫生—基础医学]