改良的人脐带间充质干细胞培养方法

目的:建立人脐带间充质干细胞(hUCMSCs)的分离和培养方法,探讨hUCMSCs的成脂成骨分化潜能。方法:用酶消化法、传统组织块法和改良组织块法从人脐带中分离间充质干细胞,流式细胞仪检测其免疫表型;用不同的培养体系诱导hUCMSCs向成骨细胞及成脂细胞分化,并对其进行鉴定。结果:改良的hUCMSCs培养方法培养细胞纯度更高,在相同的培养时间内,改良组织块法所获得的细胞数量是酶消化法的2～3倍,是传统组织块法的20～30倍,细胞呈长梭形生长;高表达CD73、CD90、CD44、CD105,不表达CD31、CD45、CD34;茜素红染色及油红O染色证实了hUCMSCs可分化为脂肪细胞和成骨细胞。结论:建立了hUCMSCs的培养新方法,证实了hUCMSCs的增殖能力和多向分化潜能,为其治疗应用提供支持。

The modified culture method of human umbilical cord mesenchymal stem cells

Objective: To establish a method to isolate and culture human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and to study hUCMSCs' differentiation potential towards osteoblasts and lipoblasts. Methods: Mesenchymal stem cells were isolated from human umbilical cord by enzyme digestion, conventional tissue adherence and modified tissue adherence. The immunophenotypes were detected by flow cytometry. The cells were induced to differentiate into osteoblasts and lipoblasts by different culture systems. Results: The modified culture method of hUCMSCs was established. The cells were of higher purity, and the primary cell number was 2-3 times as that achieved with conventional enzyme digestion method and 20-30 times as that of conventional tissue block culture. The adherent cells showed a spindle shape and were positive for CD73, CD90, CD44 and CD105, but negative for CD31, CD45 and CD34. After induction, the differentiated cells were positive for alizarin red staining and oil red O staining. Conclusion: An effectively culture method of hUCMSCs was established, and the cells can differentiate into osteoblasts and lipoblasts.