Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (40): 7435-7438.doi: 10.3969/j.issn.1673-8225.2010.40.006

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Feasibility of in vitro CM-Dil-labeled human umbilical cord mesenchymal stem cells passage tracing

Chen Li, Wu Ben-qing, Cheng Han-rong, Huang Jin-jie, Ding Lu, Song Jin-zhi   

  1. Newborn Department, Shenzhen People’s Hospital, Second College of Clinical Medicine, Jinan University, Shenzhen  518020, Guangdong Province, China
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Wu Ben-qing, Master, Chief physician, Professor, Newborn Department, Shenzhen People’s Hospital, Second College of Clinical Medicine, Jinan University, Shenzhen 518020, Guangdong Province, China wubenqing783@126.com
  • About author:Chen Li★, Studying for master’s degree, Attending physician, Newborn Department, Shenzhen People’s Hospital, Second College of Clinical Medicine, Jinan University, Shenzhen 518020, Guangdong Province, China 5823283@qq.com
  • Supported by:

    the Key Special Foundation of Medical Science of Shenzhen City, No. 20091998A03*; the Science and Technology Project of Shenzhen City in 2010, No. 201002001*

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Abstract:

BACKGROUND: The key to study biological characteristics of human umbilical cord mesenchymal stem cells (hUCMSCs) is to master their tracing method of hUCMSCs transplantation.
OBJECTIVE: To analyze the tracing feasibility of CM-Dil labeled hUCMSCs in vitro.
METHODS: hUCMSCs were isolated and cultured in vitro after enzyme digestion. Immunophenotype and cell cycle were analyzed by flow cytometry, as well as induction of the adipogenic, osteogenic differentiation of hUCMSCs were identified in vitro. The fifth passage of cells were labeled with CM-Dil, passaged and observed by fluorescence microscope in vitro.  
RESULTS AND CONCLUSION: hUCMSCs at passage 3 were strongly positive for CD44 and CD29, weakly positive for CD106, but negative for CD34 and CD40. Cells in G0/G1 phase accounted for more than 80%. Following adipogenic and osteogenic induction, oil red and alkaline phosphatase staining were positive respectively. The efficiency of CM-DiI labeling hUCMSCs was above 90%. Fluorescence intensity gradually decreased in vitro. After passaged for 8 generations, it nearly quenched. These indicated that hUCMSCs have great potential to proliferation and differentiation. The tracing method of CM-DiI labeling hUCMSCs is effective and easy.

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