论文部分内容阅读
目的:观察活体染料羧基荧光素乙酰乙酸(CFSE)标记的人羊膜间充质干细胞对四氯化碳诱导小鼠肝损伤模型的定位修复情况。方法:采用胰蛋白酶-胶原酶消化法从羊膜组织中分离间充质干细胞,通过流式细胞术和免疫荧光等方法进行鉴定。模型组按浓度为20μl/g剂量的四氯化碳和橄榄油混合液诱导小鼠肝损伤,治疗组经小鼠尾静脉注射羧基荧光素乙酰乙酸标记的人羊膜间充质干细胞约1×106个/ml。分别取模型组和细胞移植的治疗组小鼠眼球血和肝组织进行相关检测。结果:分离得到纯度较高的羊膜间充质干细胞;冰冻切片免疫荧光显示移植1周后细胞向小鼠受损肝组织定植,CFSE标记的人羊膜间充质干细胞呈绿色荧光;细胞移植后4周,与模型组比较,细胞移植组小鼠血清中天冬氨酸转移酶、丙氨酸氨基转移酶显著降低,而白蛋白明显升高(P<0.01);肝组织病理切片模型组小鼠细胞水肿,坏死灶多见,脂肪变性,可见不同程度的炎性细胞浸润;治疗组小鼠肝组织病理学改变和损伤程度有较明显改善;小鼠肝组织冰冻切片的免疫荧光显示移植4周后人羊膜间充质干细胞周围分泌血清白蛋白。结论:羧基荧光素乙酰乙酸标记的人羊膜间充质干细胞可有效改善肝组织的生理功能,为细胞定位移植治疗肝脏疾病的修复情况提供实验数据。
OBJECTIVE: To observe the localization of human amniotic membrane-derived mesenchymal stem cells labeled with carboxyfluorescein acetoacetate (CFSE) on the carbon tetrachloride-induced liver injury model in mice. Methods: Mesenchymal stem cells were isolated from amniotic tissue by trypsin-collagenase digestion and identified by flow cytometry and immunofluorescence. The mice in the model group were treated with carbon tetrachloride and olive oil at the concentration of 20μl / g, and the mice in the treatment group were injected with carboxyfluorescein acetoacetate labeled human amniotic membrane-derived mesenchymal stem cells about 1 × 106 / Ml. Were taken model group and cell transplantation in mice treated with eyeball blood and liver tissue related testing. Results: The amniotic membrane-derived mesenchymal stem cells were isolated with high purity. Immunofluorescence staining showed that the cells were implanted into the damaged liver tissue of mice one week after transplantation, and CFSE-labeled human amniotic mesenchymal stem cells showed green fluorescence. After transplantation, Compared with the model group, the serum aspartate aminotransferase and alanine aminotransferase in the cell transplantation group decreased significantly and the albumin level increased significantly (P <0.01). Compared with the model group, the mice in the liver transplantation group Cell edema and necrosis were more common, fatty degeneration, showing varying degrees of inflammatory cell infiltration; the treatment group mice liver pathology changes and the degree of injury has been significantly improved; immunofluorescence staining of mouse liver tissue frozen sections showed 4 weeks after transplantation Serum albumin is secreted by human amniotic mesenchymal stem cells. CONCLUSION: Carboxyfluorescein acetoacetate labeled human amniotic membrane-derived mesenchymal stem cells can effectively improve the physiological function of liver tissue and provide experimental data for the repair of liver diseases by cell localization and transplantation.