Chinese Journal of Tissue Engineering Research

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Growth state and adenovirus infection efficiency of human umbilical cord-derived mesenchymal stem cells in 3 different culture systems

Hong Jing-xin1, Zhang Qian-zhen2,3, Han Jun-ling1,4, Liu Hui2, Liu Jian1, Qiu Lu-gui1,4   

  1. 1Union Stem Cell & Gene Engineering Co.,Ltd., Tianjin Cord Blood Bank, Tianjin   300384, China;
    2Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University of Chinese PLA, Shanghai   200438, China;
    3Xinyuan Institute of Medicine and Biotechnology, College of Life Science, Zhejiang Sci-Tech University, Hangzhou   310018, Zhejiang Province, China;
    4Institute of Hematology, Blood Disease Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin 300020, China
  • Online:2010-01-04 Published:2010-01-04
  • Contact: Qiu Lu-gui, Chief physician, Professor, Doctoral supervisor, Union Stem Cell & Gene Engineering Co.,Ltd., Tianjin Cord Blood Bank, Tianjin 300384, China; Institute of Hematology, Blood Disease Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin 300020, China drqiu99@medmail.com.cn
  • About author:Hong Jing-xin☆, Ph.D., Assistant researcher, Union Stem Cell & Gene Engineering Co.,Ltd., Tianjin Cord Blood Bank, Tianjin 300384, China hjx78@126.com
  • Supported by:

    Special Fund for Scientific and Technological Innovation of Tianjin City, No. 08 FDZDSH03000*

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Abstract:

BACKGROUND: In vitro culture condition and culture efficiency are different in reported umbilical cord-derived mesenchymal stem cells, and lacked of unified standards. Different derived mesenchymal stem cells have different biological properties. Therefore, it is very necessary to establish a simple and high-performance culture system for umbilical cord-derived mesenchymal stem cells.
OBJECTIVE: To observe the growth state of human umbilical cord-derived mesenchymal stem cells in different culture systems in vitro and adenovirus infection efficiency.
METHODS: Mesenchymal stem cells were separated from healthy full-termed delivery fetus using collagenase digestion method and purified by adherent culture. These cells were cultured and amplified in DMEM (low glucose), MesenPRO RS™ Medium and STEMPRO® MSC SFM in vitro. The 3-5 passage mesenchymal stem cells were infected by the Ad5-EGFP, Ad5/11-EGFP, Ad5/35-EGFP as multiplicity of infection (MOI)=1, 10, 100. Viral infection and green fluorescence expression were observed at post-infection 24, 56 and 72 hours using inverted fluorescence microscope.
RESULTS AND CONCLUSION: The cell morphology in STEMPR® MSC SFM was different from other two culture system and these cells were not easy to adherent after trypsin digestion. Cell doubling time in the MesenPRO RS™ Medium was shorter than other two groups. Mesenchymal stem cells were infected by Ad5/35-EGFP with higher efficiency than other two kinds of adenovirus, but part of cells appeared apoptosis. The infection efficiency of Ad5/11-EGFP was highest. The fluorescence intensity was gradually increased with increased MOI.

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