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Objective To detect the sources of newborn rat cerebral cortex astrocytes and microglial cells, stem cell antigen expression, biological experimental basis for further in-depth study of the application of both glial cells significance. Method 1. Primary mixed glial cell cultures from neonatal rat cerebral cortex, the adhesion characteristics according to the of glial growth time difference and cells isolated by mechanical shock and differential adherence purified O-2A progenitor cells, microglial cells (microglia, MGs) and type-1 astrocytes (type 1 astrocytes, T1As), using a cell-specific antibody were identified. 2 to collect the O-2A progenitor cells seeded in 12-well culture plate and / or the packet is a PLL coverslip hydroponic solution for DMEM/F-12, 2% B27, 1% N2, 10ng/ml of PDGF, further purified O -2A progenitor cells. O-2A progenitor cells induced to differentiation of type 2 astrocytes (type 2 astrocytes, T2As) containing 20? S culture conditions, using a cell-specific antibody were identified. 3. Immunofluorescence staining method to detect the stem cell marker molecule Nestin (of nestin), stage-specific embryonic antigen -1 (SSEA-1) and proteoglycan NG2 expression in astrocytes and microglia situation. Results by mechanical shock and differential adherence, effective separation of O-2A progenitor cells and type 1 astrocytes cells, they have the typical morphological characteristics, the expression of each cell specific molecule, O-2A progenitor cells expression A2B5 1 astrocytes expressed GFAP. Mechanical shock and differential adherence, effective separation of purified microglia, they have the typical morphological characteristics, and expression of cell-specific molecule of OX42 separate O-2A progenitor cells after PDGF proliferation without differentiation thus purified O-2A progenitor cells. 4.O-2A progenitor cells in serum-free culture medium, rapidly differentiate into mature oligodendrocytes; containing up to 15% -20? S hydroponic solution growth, differentiation, type 2 astrocytes, specific antigen expression A2B5 / GFAP. 5. Immunofluorescence staining revealed: the differentiation and maturation of type 2 astrocytes express nestin, SSEA-1 and NG2, microglial cells express nestin, SSEA-1 and NG2 1 astrocytes did not express stem cell antigen markers. Conclusions of the neural stem cell marker nestin, differences SSEA-1 and NG2 expression in glial cells. Type 2 astrocytes and microglial cells express nestin and SSEA-1 and NG2, has the certain stem cell antigen expression characteristics, combined with I Laboratory has been to study the expression of prompt, type 2 astrocytes and small glial cell biology of neural stem cell-like characteristics, can be used as cell sources used in the treatment of central nervous system diseases.
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