Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (27): 4955-4959.doi: 10.3969/j.issn.1673-8225.2010.27.004

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Monolayer coculture of rabbit chondrocytes and bone marrow stromal cells: Screening of culture proportion

Sun Lei1, Qi Hui1, Chen Lei1, Chen Hai-yan1, Jiang Jian1, Tao Jian-feng1, Jie Yong-sheng1, Gao Xin-sheng1, Peter I. Lelkes2   

  1. 1Beijing Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Beijing   100035, China;
    2School of Biomedical Engineering, Drexel University, United States
  • Online:2010-07-02 Published:2010-07-02
  • About author:Sun Lei, Doctor, Associate investigator, Beijing Institute of Traumatology and Orthopaedics, Beijing Jishuitan Hospital, Beijing 100035, China dr.sunlei2009@gmail.com
  • Supported by:

    the Science and Technology New Star Project of Beijing City, No. H013610310113*

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Abstract:

BACKGROUND: Bone marrow stromal cells (BMSCs) can differentiate into chondrocytes under a certain condition, but there are lack of studies concerning monolayer coculture of chondrocytes and BMSCs at present.

OBJECTIVE: To identify percentage choice of coculture of BMSCs and chondrocytes, cell proliferation activity and cartilage specific protein expression rule, and to find the best time and subculture frequency during cell transplantation.

METHODS: Chondrocytes and BMSCs were isolated, cultured, and divided into 5 groups following passage: chondrocyte group, coculture (chondrocytes and BMSCs at 7: 3, 5: 5, 3: 7) groups, and BMSC group. At generation 4(G4), cell morphology was observed at different passages in each group under an inverted phase contrast microscope. MTT assay was performed for testing the cell viability. The proteoglycan expression was detected by toluidine blue and alcian blue staining. Type II collagen expression was determined by immunocytochemical method.

RESULTS AND CONCLUSION: Cells showed normal appearance in all the coculture groups. From G1-G3, cells had good viability, which was decreased at G4. Proteoglycan (detected by toluidine blue and alcian blue staining) and type II collagen expression were highly positively determined in the coculture groups during the second and third generations (G2 and G3). Synthetic each index showed that each index was best in the chondrocytes and BMSCs at 5: 5, 3: 7 groups. Results displayed that monolayer coculture of chondrocytes and BMSCs kept chondrocyte morphology and protein expression property. The proportion of chondrocytes and BMSCs at 5: 5 or 3: 7 was optimal for cell transplantation.

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