Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (6): 1027-1030.doi: 10.3969/j.issn.1673-8225.2010.06.016

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Four kinds of extracellular matrixes support the growth of human embryonic stem cells: Effect differences

Hu Zhi-xing1,2, Luo Min2,3, Zhou Yi-ping2,3, Liang Dao-ming4   

  1. 1Department of Pharmacology, Kunming Medical University, Kunming   650031, Yunnan Province, China;
    2Biomedical Engineering Research Center, Kunming Medical University, Kunming   650031, Yunnan Province, China; 
    3Yunnan Provincial Pharmacological Key Laboratory of Natural Drug, Kunming Medical University, Kunming   650031, Yunnan Province, China;
    4Second Affiliated Hospital, Kunming Medical University, Kunming 650031, Yunnan Province, China
  • Online:2010-02-05 Published:2010-02-05
  • Contact: Liang Dao-ming, Doctor, Associate chief physician, Second Affiliated Hospital, Kunming Medical University, Kunming 650031, Yunnan Province, China liangdaoming@yahoo.com.cn
  • About author:Hu Zhi-xing, Doctor, Lecturer, Department of Pharmacology, Kunming Medical University, Kunming 650031, Yunnan Province, China; Biomedical Engineering Research Center, Kunming Medical University, Kunming 650031, Yunnan Province, China xingzhihu@163.com
  • Supported by:

    Special Research Foundation of Department of Science and Technology of Yunnan Province-Kunming Medical University, No.2007C0012R, 2007C0044R**;
    Natural Science Foundation of Yunnan Province, No.2005C0013R*

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Abstract:

BACKGROUND: Extracellular matrixes play important roles in the maintenance of undifferentiated state and self-renewal of human embryonic stem cells (hESCs). Establishment of a defined serum- and feeder-free culture system is a prerequisite for the application of hESCs in cell-replacement therapy.

OBJECTIVE: The present study was designed to compare the effects of four kinds of extracellular matrixes on the support the growth of hESCs.

METHODS: We set four groups using conventionally recovered hESCs BGO2. BG02 was cultured on Matrigel-, laminin-, fibronectin-, or collagen-coated plates in serum-free medium containing basic fibroblast growth factor (bFGF), transforming growth factor beta 1(TGFβ1), Insulin-Transferrin-Selenium (ITS). The attachment rate and differentiated rate of BG02 clumps in various conditions were measured. The proportions of Oct-4 and Nanog positive cells in various groups were assessed by flow cytometry. The extracellular matrix receptor expression was analyzed by RT-PCR.

RESULTS AND CONCLUSION: The attachment rates of BG02 cell clumps in Matrigel and laminin groups were significantly higher than that of fibronectin and collagen groups (P < 0.05), but differentiated rate was significantly reduced (P < 0.05). The percentage of Oct-4+ and Nanog+ BG02 cells in Matrigel and laminin groups were higher than that of fibronectin and collagen groups (P < 0.05). No significant difference in each index was determined between the fibronectin and collagen groups (P > 0.05). BG02 cells in Matrigel and laminin groups expressed higher levels of integrin α5, integrin α6 and integrin β1 mRNA than that in fibronectin and collagen groups. Above-mentioned results indicated that Matrigel and laminin could support the proliferation of BG02 cells well, while fibronectin and collagen could not support the proliferation of BG02 cells. These variations might be attributed to the activation of integrin subunits in BG02 cells.

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