Chinese Journal of Tissue Engineering Research ›› 2013, Vol. 17 ›› Issue (40): 7034-7039.doi: 10.3969/j.issn.2095-4344.2013.40.003

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Differentiation of human umbilical cord blood derived-mesenchymal stem cells into chondrocytes induced by supernatants of human chondrocytes

Zhou Feng, Wang Ying-zhen, Zhang Hai-ning, Lü Cheng-yu, Xu Zong-yao   

  1. Qingdao University Medical School, Qingdao  266003, Shandong Province, China
  • Online:2013-10-01 Published:2013-10-31
  • Contact: Zhang Hai-ning, Associate chief physician, Qingdao University Medical School, Qingdao 266003, Shandong Province, China
  • About author:Zhou Feng★, Master, Qingdao University Medical School, Qingdao 266003, Shandong Province, China yuyenongcha@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 81171774*, 81272056*

Abstract:

BACKGROUND: Human umbilical cord blood derived-mesenchymal stem cells have been used as seed cells to repair cartilage defects. Their application has been greatly limited by high cost and sufficient amount although they could be cultured in induced medium or induction factors.
OBJECTIVE: To verity the feasibility to induce human umbilical cord blood derived-mesenchymal stem cells to differentiate into the chondrocytes using supernatants of human chondrocytes.
METHODS: Human umbilical cord blood derived-mesenchymal stem cells were separated with density gradient centrifugation and adherent method from human umbilical cord blood. Surface markers of human umbilical cord blood derived-mesenchymal stem cells were identified by flow cytometry. Human hyaline cartilage was dissected, digested, and chondrocytes were cultured in vitro. The human umbilical cord blood derived-mesenchymal stem cells were cultured with supernatant from chondrocytes. After 2 weeks, the changes of phenotype of the cells were identified, and type II collagen expression was detected using immunohistochemistry.
RESULTS AND CONCLUSION: Human umbilical cord blood derived-mesenchymal stem cells were isolated from umbilical cord blood using density gradient centrifugation and adherent method. Flow cytometry demonstrated that the human umbilical cord blood derived-mesenchymal stem cells expressed CD90 and CD105 positively, failed to express CD34 or CD45. After 2 weeks of induction, human umbilical cord blood   
derived-mesenchymal stem cells turned to polygon and round shaped. Immunohistochemistry staining showed some human umbilical cord blood derived-mesenchymal stem cells expressed type II collagen. Results indicate that culture medium of chondrocytes can induce human umbilical cord blood derived-mesenchymal stem cells to express type II collagen, and differentiate into the chondrocytes.

Key words: stem cells, chondrocytes, collagen type II, centrifugation, density gradient

CLC Number: