中国组织工程研究Issue(23)：4196-4203,8.DOI:10.3969/j.issn.2095-4344.2013.23.003
共培养诱导人脐血间充质干细胞向软骨细胞分化*★
Chondrogenic differentiation of co-cultured human umbilical cord blood-derived mesenchymal stem cells
摘要
Abstract
BACKGROUND: Human umbilical cord blood-derived mesenchymal stem cel s can be induced through the co-culture to differentiate into other cel s; however, it is difficult to control the proportions of two kinds of cel s for maximizing the efficiency of obtaining target cel s. OBJECTIVE: To investigate chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cel s by co-culture with rabbit chondrocytes at different ratios. METHODS: Human umbilical cord blood-derived mesenchymal stem cel s were resuscitated and cultured, and identified by flow cytometry. Then human umbilical cord blood-derived mesenchymal stem cel s were co-cultured with rabbit chondrocytes at 2:1 or 3:1 ratio, and induced with insulin-like growth factor-1. After 14 days in co-culture, the cel RNA and protein were extracted, aggrecan mRNA and col agen type Ⅱ mRNA expression levels were detected by real-time quantitative-polymerase chain reaction, and aggrecan and col agen type Ⅱ protein expression levels were detected by western blot assay. RESULTS AND CONCLUSION: After human umbilical cord blood-derived mesenchymal stem cel s were co-cultured with rabbit chondrocytes for 14 days, the aggrecan mRNA and col agen type 2 mRNA expression in the co-cultured cel s was significantly higher than that in the cel s only induced with insulin-like growth factor-1. Under the same ratio of human umbilical cord blood-derived mesenchymal stem cel s and rabbit chondrocytes, insulin-like growth factor-1 increased the aggrecan and col agen type Ⅱ protein and mRNA expressions. A 3:1 co-culture ratio al owed the increment in the col agen type Ⅱ mRNA and protein expression, as wel as aggrecan protein expression; while a 2:1 co-culture ratio upregulated aggrecan mRNA expression. Experimental findings indicate that, human chondrocytes can be successful y induced by co-culture of human umbilical cord blood-derived mesenchymal stem cel s and rabbit chondrocytes, and 3:1 co-culture ratio can obtain more chondrocytes.关键词
干细胞/脐带脐血来源干细胞/软骨细胞/共培养/胰岛素样生长因子/Ⅱ型胶原/聚集蛋白多糖/软骨缺损/软骨组织工程/种子细胞/其他基金Key words
stem cel s/umbilical cord/umbilical cord blood stem cel s/chondrocytes/co-culture/insulin-like growth factor-1/col agen type Ⅱ/aggrecan/cartilage defects/cartilage tissue engineering/seed cel s/other grants-supported paper分类
医药卫生引用本文复制引用
郑朋飞,陈雷,董展,蒋立,鞠黎,王儒法,楼跃..共培养诱导人脐血间充质干细胞向软骨细胞分化*★[J].中国组织工程研究,2013,(23):4196-4203,8.基金项目
2010南京市卫生局课题(QYK10163)。 （QYK10163）
