›› 2012, Vol. 32 ›› Issue (8): 1106-.doi: 10.3969/j.issn.1674-8115.2012.08.031

• Technique and method • Previous Articles    

Isolation, culture and identification of rat articular cartilage stem cells

TONG Wen-xue1, XIANG Sheng-nan1, ZHANG Ning1, DAI Ke-rong1,2, ZHANG Xiao-ling1,2   

  1. 1.The Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Jiaotong University School of Medicine &|Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China；2.Shanghai Key Laboratory of Orthopaedic Implant, Department of Orthopaedics, the Ninth People´s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2012-08-28 Published:2012-08-29
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 11QH1401600；Shanghai Education Committee Foundation, 10SG22, J50206

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Abstract:

Objective To isolate and culture rat articular cartilage stem cells (ACSCs), and identify their characteristics. Methods ACSCs were isolated from rat articular cartilages by fibronectin-conglutination assay. The expression of positive stem cell surface markers (CD90 and CD44) and negative stem cell surface markers (CD45, CD31 and CD34) in ACSCs was detected by flow cytometry. The single cell colony-forming efficiency of ACSCs was determined by single cell colony-forming assay. The multidirectional differential potential of ACSCs was identified by osteogenesis, adipogenesis and chondrogensis experiment. Results ACSCs were successfully isolated. CD44 and CD90 were highly expressed in ACSCs, while CD31, CD34 and CD45 were not expressed in ACSCs. Seven days after culture, single ACSC formed single cell colony containing more than 32 cells. ACSCs demonstrated a high osteogenesis, adipogenesis and chondrogenesis potential. Conclusion ACSCs are present in rat articular cartilages, and ACSCs have typical characteristics of stem cells.

Key words: articular cartilage stem cells, cartilage, fibronectin, cartilage regeneration