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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 whether fetal connective tissue can induce bone marrow mesenchymal stem cells (BMSCs) to differentiate into epithelial cells. Methods Tissue pieces containing fetal intestinal mucosa and submucosa were prepared and enzymatically digested enterocytes. The 3rd generation BMSCs labeled with DAPI were planted on the submucosa of the tissue block with and without A and B EGF groups. C and D control groups of BMSCs were also established. In vitro cultured to 12 days, doing light microscope, CLSM morphology, histochemistry and immunohistochemistry. Results ① Fluorescence microscopy showed that there were DAPI fluorescent labeled cells on the surface of the tissue, HE staining showed that the cells were epithelial-like. ② Immunohistochemical staining showed that the surface cells of group A and B and the cells of group C could express CK and CK20, while group D was negative; EGF of intestinal tissue was expressed. CLSM under the visible surface of the tissue block blue DAPI and red CK or green EMA double fluorescent positive cells. ③ PAS response showed a basal membrane-like structure between the cells and connective tissue. Conclusion Connective tissue of intestinal wall can induce BMSCs to differentiate into epithelial cells. Endogenous EGF may be one of the inducing factors.