Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (10): 1721-1728.doi: 10.3969/j.issn.1673-8225.2010.10.003

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Coculture of human umbilical cord mesenchymal stem cells from Wharton’s jelly and brain tumor stem cells

Tian Yi1,2, Guan Fang-xia3, Hu Xiang4,5, Yang Bo1,2, Du Ying6, Zhou Chang-hui3, Ba Yun-tao1, Gu Chen-xi7, Lei Ning-jing3, Wang Xiao-wei8   

  1. 1Department of Neurosurgery, First Affiliated Hospital, Zhengzhou University, Zhengzhou   450052, Henan Province, China;
    2Institute of Clinical Medicine, Zhengzhou University, Zhengzhou   450052, Henan Province, China;
    3Department of Bioengineering, 7Department of Clinical Medicine, Zhengzhou University, Zhengzhou   450001, Henan Province, China;
    4Jiangsu Public Technology Service Platform of Stem Cells and Biotherapy, Taizhou   225300, Jiangsu Province, China;
    5Shenzhen Beike Cell Engineering Institute, Shenzhen   518000, Guangdong Province, China;
    6Department of Microbiology and Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou   450052, Henan Province, China;
    8College of Life Science, Hunan Normal University, Changsha   410081, Hunan Province, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: Yang Bo, Master, Professor, Doctoral supervisor, Department of Neurosurgery, First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, Henan Province, China; Institute of Clinical Medicine, Zhengzhou University, Zhengzhou 450052, Henan Province, China yangbo96@126.com
  • About author:Tian Yi, Studying for Master’s degree, Physician, Department of Neurosurgery, First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, Henan Province, China; Institute of Clinical Medicine, Zhengzhou University, Zhengzhou 450052, Henan Province, China ethan43@qq.com Guan Fang-xia, Doctor, Professor, Department of Bioengineering, Zhengzhou University, Zhengzhou 450001, Henan Province, China guanfangxia@126.com Tian Yi and Guan Fang-xia contributed equally to this article.
  • Supported by:

    the Zhengzhou University “211 Project-Phase Ⅲ”- the Basic and Clinical Research of Stem Cells*; Jiangsu Public Technology Service Platform Development Program of Stem Cells and Biotherapy, No. BM2008146*

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Abstract:

BACKGROUND: Human mesenchymal stem cells derived from Wharton’s jelly (WJCs) display the characteristics of MSCs as defined by the International Society for Cellular Therapy. They can be differentiated into bone, cartilage, adipose, muscle, and neural cells. They can also support the expansion of other stem cells, be well-tolerated by the immune system, and have the ability to home to tumors.

OBJECTIVE: To investigate biological changes of WJCs and brain tumor stem cells (BTSCs) co-cultured in vitro.

METHODS: WJCs cultured by situ cultivation and BTSCs used enzyme digestion way respectively, and gathering the 3rd passage of WJCs though subculturing as well as BTSCs. Two kinds of cells co-cultured in 24-well plates in serum-free medium (SFM) without any growth factor. 3 and 7 days after co-cultured respectively, CD133 expression of suspension cells in the 24-well plates were identified by flow cytometry, and immunofluorescence was performed for Nestin and glial fibrillary acidic protein (GFAP) expression of adherent cells. Co-culture supernatant (CCS) re-suspended 3rd passage of BTSCs and cultured into 96-well plates at day 3, which were used to determine the difference in cell growth curve in both groups using a microplate reader.

RESULTS AND CONCLUSION: With the cocultivation days increasing, the phenomenon that tumor sphere cells began to be decomposed, adherent and differentiated observed by an inverted microscope. BTSCs in the co-cultured group expressed GFAP and Nestin when adherent and differentiated. The higher degree of malignant brain tumor tissue used in culturing BTSCs was, the higher expression of CD133 in BTSCs was. CD133+ in BTSCs declined when co-cultured with WJCs. Growth curve of brain tumor stem cells cultured in CCS compared with in SFM at day 3, which indicates that the proliferation of BTSCs inhibited obviously. Results indicated that CD133+ expression and proliferative capacity of BTSCs went down and BTSCs underwent differentiation during the co-culture in vitro.

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