Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (41): 7625-7630.doi: 10.3969/j.issn.2095-4344.2012.41.006

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Induced differentiation of human umbilical cord mesenchymal stem cells into chondrocytes in two kinds of culture systems

Liu Jing1, Xu Chao1, Yu Li1, Luo Er-mei1, Zheng Yin-li1, Wu Ying-hua1, Chen Jian-ye1, Zhou Zi-peng1, Tang Ming-qiao2   

  1. 1Department of Biochemistry, School of Medicine, Jinan University, Guangzhou 510632, Guangdong Province, China
    2Department of Mechanics and Civil Engineering, College of Technology, Jinan University, Guangzhou 510632, Guangdong Province, China
  • Received:2012-01-03 Revised:2012-02-24 Online:2012-10-07 Published:2012-10-07
  • Contact: Tang Ming-qiao Lecturer, Department of Mechanics and Civil Engineering, College of Technology, Jinan University, Guangzhou 510632, Guangdong Province, China doctoryuli@yahoo.com.cn tmqtmq@jnu.edu.cn
  • About author:Liu Jing★, Studying for master’s degree, Department of Biochemistry, School of Medicine, Jinan University, Guangzhou 510632, Guangdong Province, China liujing-87424@163.com

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Abstract:

BACKGROUND: The methods of inducing mesenchymal stem cells to differentiate into chondrocytes under non-scaffold condition include high-density pellet culture, in vitro monolayer culture and co-culture with chondrocytes. High-density pellet culture and in vitro monolayer culture are widely concerned due to simple operation.
OBJECTIVE: To find a better method of inducing human umbilical cord mesenchymal stem cells into chondrocytes in non-scaffold culture in vitro.
METHODS: Human umbilical cord mesenchymal stem cells were isolated and cultured by tissue block method, and cell surface antigens were identified by flow cytometry. Human umbilical cord mesenchymal stem cells were cultured with chondrogenic media and stained with alcian blue. The production of matrix was estimated from the determination of hydroxyproline content using alcian blue method. Expressions of glycosaminoglycan, type Ⅱ collagen and Sox-9 were assayed by real-time fluorescence quantitative PCR.
RESULTS AND CONCLUSION: The phenotype of cultured human umbilical cord mesenchymal stem cells was CD105+/CD29+/CD44+/ CD31-/CD34-/CD40-/CD45-/HLA-DR-. Human umbilical cord mesenchymal stem cells weakly expressed chondrocyte markers, which strongly expressed glycosaminoglycan and type Ⅱ collagen after chondgenic induction, and the expressions were higher in cells cultured in pellet culture system than in monolayer culture system. Real-time quantitative PCR results demonstrated that cells expressed higher level of glycosaminoglycan, type Ⅱ collagen and Sox-9 after induction than that before induction, and the expression levels were higher in cells cultured in pellet culture system than in monolayer culture system. The results revealed that human umbilical cord mesenchymal stem cells cultured in pellet culture system are more conducive to differentiate into chondrocytes than those cultured in monolayer culture system

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