Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (23): 4222-4226.doi: 10.3969/j.issn.1673-8225.2010.23.010

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Effect of microglia on differentiation from in vitro cultured neural stem cell into cholinergic neurons

Jin Yu-ling1, Liu Qie1, Liu Jing-jing2, Li Xin1, Wang Li-hua1   

  1. 1First Department of Neurology, First Hospital Affiliated to Jiamusi University, Jiamusi   154007, Heilongjiang Province, China;
    2Department of Neurology, General Hospital of Jixi Coal Mining Bureau, Jiamusi   158120, Heilongjiang Province, China
  • Online:2010-06-04 Published:2010-06-04
  • About author:Jin Yu-ling, Associate chief physician, Professor, Master’s supervisor, First Department of Neurology, First Hospital Affiliated to Jiamusi University, Jiamusi 154007, Heilongjiang Province, China jyl2017@sina.com
  • Supported by:

    the Natural Science Foundation of Heilongjiang Province, No. D200859*

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Abstract:

BACKGROUND: Orient differentiation of neural stem cells depends on concentration of microglia. A small concentration can not play an effect to a certain degree; and a large concentration might cause cytotoxicity.

OBJECTIVE: To observe the effect of varying concentrations of microglia on differentiation from in vitro cultured neural stem cells into neurons.

METHODS: Newborn Wistar rats within 24 and 48 hours were used for primary culture of neural stem cells and microglia, which were then identified by immunohistochemistry. Neural stem cells and microglia were co-inoculated in 6-well plates, at the density of 10:1, 4:1, 1:1, 1:4, and 1:10, with 6 wells at each density. Neural stem cells alone were considered as control group and incubated for days 3, 7 and 14 to observe the cell growth.

RESULTS AND CONCLUSION: Primary culture of neural stem cells gathered like a ball and grew in a suspending manner. Nestin staining was positive, but nuclei were not stained. Microglia grew in an adherent manner, with strong refractive index. Most of the cells showed short branch-like protrusions. Microglia-specific marker antibodies CD11b/c staining showed that the cell purity was above 80%. Compared with control group, neural stem cells and microglia cells were co-cultured at a density of 10:1, 4:1, 1:1, 1:4, and 1:1. ChAT-positive cells were significantly increased. If neural stem cells and microglia were co-cultured at the density of 4:1, the increasing was significantly greater than other co-cultured statuses (P < 0.05), and the cell growth peaked at 7 days. The microglia could promote differentiation from neural stem cells into cholinergic neurons, and the co-culture of neural stem cells and microglia at a density of 4:1 achieved the best effect at 7 days.

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