Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (32): 5949-5951.doi: 10.3969/j.issn.1673-8225.2010.32.013

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Differentiation of human placenta-derived mesenchymal stem cells into cholinergic neuron-like cells in vitro 

Jing Xu-dong, He Dong-mei, Zhang Yuan, Chen Li   

  1. Institute of Hematology, Medical College of Jinan University, Guangzhou  510632, Guangdong Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Zhang Yuan, Doctor, Researcher, Institute of Hematology, Medical College of Jinan University, Guangzhou 510632, Guangdong Province, China tzyuan@jnu.edu.cn
  • About author:Jing Xu-dong★, Master, Attending physician, Institute of Hematology, Medical College of Jinan University, Guangzhou 510632, Guangdong Province, China thedm@jnu.edu.cn
  • Supported by:

    the Key Subjects of the State Council Sponsored by Overseas Chinese, No. 51205002*

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Abstract:

BACKGROUND: Human placenta may contain similar mesenchymal stem cells (MSCs), which can be differentiated into cholinergic neuron-like cells.
OBJECTIVES: To explore the differentiation of MSCs from human placenta into cholinergic neuron-like cells in vitro.
METHODS: MSCs from human placenta were isolated and purified by cell culture. The fourth passage of MSCs was incubated with fetal bovine serum+basic fibroblast growth factor (bFGF) for 2 days, and divided into 3 groups: groupⅠ, MSCs were incubated with DMEM/F12 complemented with bFGF, 2-mercaptoethanol, retinoic acid and nerve growth factor for 19 days. GroupⅡ, based on Group Ⅰ, epidermal growth factor and heparin were added into the culture system for 19 days. In the negative control group, cells were cultured by DMEM/F12 containing fetal bovine serum.
RESULTS AND CONCLUSION: The adhesive cells gradually formed flat monolayer cells, parallel arranged or whirlpool grew, with uniform morphology after 2 passaged. The fourth passage of MSCs from human placenta was strongly positive for CD44, CD29, but negative for CD34, CD45, CD106 and HLA-DR. After 2 weeks of culture, nestin, chat mRNA could be expressed. The positive rate of nestin, Ache and chat mRNA was higher in the group Ⅰ and group Ⅱ than that of the negative control group (P < 0.05). Compared to the negative control group, the content of acetylcholine in supernate was obviously increased in the group Ⅰ and group Ⅱ (P < 0.05), especially more in the group Ⅰ (P < 0.05). The results demonstrated that MSCs from human placeta can differentiate into cholinergic neuron-like cells, which are able to synthesis, release and disintegrate acetylcholine.

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