Chinese Journal of Tissue Engineering Research ›› 2013, Vol. 17 ›› Issue (10): 1711-1718.doi: 10.3969/j.issn.2095-4344.2013.10.001

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A combined use of growth factors promotes ligament cell-like cell differentiation of bone marrow mesenchymal stem cells

Sun Fang-fei, Zhang Chun-li, Li Xiao-jian, Han Xian-wei, Li Guang-zheng   

  1. Department of Sport Injury, Xijing Orthopedic Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China
  • Received:2012-09-27 Revised:2013-01-16 Online:2013-03-05 Published:2013-03-05
  • Contact: Zhang Chun-li, M.D., Chief physician, Master’s supervisor, Department of Sport Injury, Xijing Orthopedic Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China zhangcl816@sohu.com
  • About author:Sun Fang-fei, Department of Sport Injury, Xijing Orthopedic Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China 912857695@qq.com

Abstract:

BACKGROUND: A key to construction of tissue-engineered ligament is to induce bone marrow mesenchymal stem cells into ligament cells like cells using growth factors and to promote the differentiated cells to secrete collagen protein. Growth factors basic fibroblast growth factor and transforming growth factor β1 play an important role in oriented differentiation of cells.
OBJECTIVE: To in vitro induce rabbit bone marrow mesenchymal stem cells to differentiate into ligament cell-like cells using basic fibroblast growth factors and transforming growth factor β1 and investigate the biological characteristics of the ligament cell-like cells.
METHODS: Bone marrow mesenchymal stem cells were harvested and cultured. Then these cells were induced using 25 μg/L basic fibroblast growth factors and 10 μg/L transforming growth factor β1. Four groups were designated: namely blank control group, transforming growth factor β1 group, basic fibroblast growth factor group and transforming growth factor β1 + basic fibroblast growth factor group (combination group). Effects of both growth factors on the growth and morphology of bone marrow mesnchymal stem cells were investigated. Cell growth curve was drawn using MTT assay. The mount of collagen secreted by bone marrow mesenchymal stem cells was quantified using Picro-Sirius red staining. After BrdU fluorescent staining, bone marrow mesenchymal stem cells induced by transforming growth factor β1 combined with basic fibroblast growth factor were transplanted onto acellular dermal matrix to determine cell proliferation and distribution. Comparisons were made between the combination group and blank control group.
RESULTS AND CONCLUSION: Cell morphology in the combination group, as well as cell proliferation rate and collagen secretion amount, was superior to that in the blank control group, transforming growth factor β1 group, or basic fibroblast growth factor group. These findings suggest that transforming growth factor β1 combined with basic fibroblast growth factor can promote bone marrow mesenchymal stem cells to differentiate into ligament cell-like cells, which exhibits important significance for construction of tissue-engineered anterior cruciate ligament.

Key words: stem cells, bone marrow-derived stem cells, bone marrow mesenchymal stem cells, growth factor, anterior cruciate ligament, basic fibroblast growth factor, transforming growth factor, acellular dermal matrix, provincial grants-supported paper, stem cell photographs-containing paper

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