两性霉素B处理人脐带间充质干细胞的培养与分化

背景：建立和完善避免真菌污染的方法，有效降低采集脐带时分离培养的污染率，以便获得优质高效的脐带间充质干细胞。目的：探讨两性霉素B对脐带间充质干细胞的生长状态及分化潜能的影响。方法：采用胶原酶消化法从正常顺产足月新生儿脐带中分离出间充质干细胞，经两性霉素 B 处理，用MesenPRO RS？培养基进行体外扩增培养。取对数生长期的第3代脐带间充质干细胞，分析其形态、增殖方式和免疫表型，在体外诱导其向成骨和脂肪细胞分化。结果与结论：经两性霉素B处理后，在体外仍能成功分离培养出脐带间充质干细胞。流式细胞术结果显示，人脐带间充质干细胞仍强表达CD44，CD105，CD73，CD90，阴性表达HLA-DR，CD29，CD31，CD34。两性霉素B处理后的人脐带间充质干细胞经体外诱导能向脂肪和成骨细胞分化。

Cultivation and differentiation of human umbilical cord-derived mesenchymal stem cells treated with amphotericin B

High-quality and efficient umbilical cord-derived mesenchymal stem cells could be obtained by establishing and improving the method to avoid fungal contamination and by effectively decreasing pol ution rate of isolated culture during umbilical cord col ection. OBJECTIVE：To explore the effects of amphotericin B on growth and differentiation potential of umbilical cord-derived mesenchymal stem cells. METHODS：Umbilical cord-derived mesenchymal stem cells were separated from healthy ful-termed delivery fetus using col agenase digestion method and treated with amphotericin B. Subsequently, umbilical cord-derived mesenchymal stem cells were amplified and cultured in vitro with MesenPRO RSTM medium. The third passage of umbilical cord-derived mesenchymal stem cells in logarithmic phase was obtained to analyze their morphology, proliferation and immunophenotype, and induced to differentiate into osteoblasts and adipocytes in vitro. RESULTS AND CONCLUSION：After treatment with amphotericin B, umbilical cord-derived mesenchymal stem cells were successful y isolated and cultured in vitro. Flow cytometry results revealed that human umbilical cord-derived mesenchymal stem cells strongly expressed CD44, CD105 and CD73, CD90, and negatively expressed HLA-DR, CD29, CD31, and CD34. Amphotericin B-treated human umbilical cord-derived mesenchymal stem cells can stil differentiate into adipocytes and osteoblasts in vitro.