摘要
目的观察骨髓移植后供体骨髓来源干细胞(bone marrowderived stem cells,BMDSCs)是否能够在受体肠上皮中定植并分化为肠上皮细胞。方法使用密度为1.073 g/mL的Percoll分离GFP转基因小鼠的骨髓有核细胞。利用60Co放射源对C57BL/6小鼠进行10 Gy致死剂量照射,照射后4 h经尾静脉移植绿色荧光蛋白(green fluorescence protein,GFP)转基因小鼠骨髓细胞建立嵌合小鼠模型,移植后1月和6月取小肠标本石蜡包埋。使用GFP抗体鉴定GFP阳性供体细胞在小肠上皮的表达情况,并使用GFP抗体和细胞角蛋白(PCK)抗体进行双标以判断BMDSCs是否分化上皮细胞,同时结合绒毛蛋白(Villin)和GFP的免疫荧光双标及连续切片染色肠道特异转录因子CDX2和GFP来分析骨髓来源细胞是否分化为肠道功能性上皮细胞。结果供体骨髓来源细胞在嵌合鼠的骨髓和小肠上皮层中均有定植,且小肠上皮中GFP阳性细胞能够同时表达PCK、Villin和CDX2。结论供体骨髓来源干细胞可以在骨髓嵌合小鼠的肠上皮定植,并分化为具有正常功能的小肠上皮细胞。
Objective To observe whether bone marrow derived stem cells （BMDSCs） can implant into the recipient＇s intestinal epithelium and differentiate into functional intestinal epithelial cells after bone marrow transplantation. Methods Bone marrow karyocytes of GFP transgene mice were isolated by Percoll with a density of 1. 073 g/mL. Normal C57BL/6 mice were suffered from ^60Co whole body irradiation （WBI） with a lethal dose of 10 Gy. Four hours later, they received bone marrow transplantation by injection through the caudal vein to form bone marrow chimeric model. Af- ter 1 month and 6 months, small intestine was fixed in neutral formaldehyde and embedded in paraffin wax. The implantation of donor＇ s BMDSCs in the intestinal epithelium was determined with GFP antibody. And double staining of GFP and pan-cytokeratin was employed to determine whether those BMDSCs had differentiated into epithelial cells. In addition, detection of Villion with GFP and serial section staining of intestinal specific transcription factor CDX2 and GFP were used to analyze whether BMDSCs could differentiate into intestinal specific functional epithelial cells. Results GFP-positive donor＇s bone marrow derived cells could implant into recipient＇ s bone marrow and small intestinal epithelium, and these cells in intestinal epithelium could co-express GFP and PCK. The positive cells also expressed Villin and CDX2. Conclusion Donor＇ s BMDSCs can implant into the intestinal epithelium of chimeric mice and they can play as normal functional intestinal epithelial cells.
出处
《胃肠病学和肝病学杂志》
CAS
2008年第8期635-638,共4页
Chinese Journal of Gastroenterology and Hepatology
基金
国家重点基础研究发展规划项目("973"项目)(2005CB522605)
国家自然科学基金(30400247)
关键词
骨髓来源干细胞
放射损伤
肠上皮细胞
转分化
BMDSCs
Irradiated injury
Intestinal epithelial cells
Transdifferentiation
