|
|
Optimization on the cultivation method in vitro of cerebral astrocytus from mice |
LIU Xiao-mei1, ZHAO Dan2, PANG Rong-rong2, SHAN Kai2, LI Yan2, WANG Ying-wei2 |
1.Department of Microbiology and Immunology, Xuzhou Medical College, Xuzhou Jiangsu 221002; 2.Department of Microbiology and Immunology, Nanjing Medical University, Nanjing Jiangsu 210029, China |
|
|
Abstract Objective: To optimize the cultivation method of cerebral astrocytes from mice in vitro, in which develop an experimental basis for the functional study of astrocytus. Methods: Cerebral cortex was obtained from 0 to 1d C57BL/6 mice and cell suspension was prepared by digestion with trypsin digestion and mechanical dissociation. After differential attachments, primary cells were cultivated for 24 h, so the fresh culture medium instead of the old was added softly. After then, the cell flask was washed shakely before the old culture medium was replaced with the fresh medium every two or three days. After three passages, astrocytus in the culture medium were stained with anti-glial fibrillary acidic protein (GFAP) antibody using immunocytochemical or immunofluorescence staining. Results: After primary cell culture and subculture, the purified astrocytes had the typical shapes with a purity beyond 95%. Conclusion: A reproducible and simple cultivation method in vitro of astrocytus was established, in which high purity astrocytes were obtained.
|
Received: 29 September 2012
|
|
|
|
[1]Haydon PG,Carmignoto G.Astrocyte control of synaptic transmission and neurovascular coupling[J]. Physiol Rev, 2006, 86(3): 1009-1031.[2] Verkhratsky A, Steinh user C. Ion channels in glial cells[J]. Brain Res Brain Res Rev, 2000, 32(2/3): 380-412.[3] Iadecola C, Nedergaard M. Glial regulation of the cerebral microvasculature[J]. Nat Neurosci, 2007, 10(11): 1369-1376.[4] Dong Y, Benveniste EN. Immune function of astrocytes[J]. Glia, 2001, 36(2):180-190.[5] Sheng J, Yang S, Xu L, et al. Bystin as a novel marker for reactive astrocytes in the adult rat brain following injury[J]. Eur J Neurosci, 2004, 20(4): 873-884.[6] Girardi E, Ramos AJ, Vanore G, et al. Astrocytic response in hippocampus and brain cortex in an experimental epilepsy model\[J\]. Neurochem Res, 2004, 29(2) :371-377.[7] Williams A, Piaton G, Lubetzki C. Astrocytesfriends or foes in multiple sclerosis?[J]. Glia, 2007, 55(13): 1300-1312.[8] Li C, Zhao R, Gao K, et al. Astrocytes: implications for neuroinflammatory pathogenesis of Alzheimer's disease[J]. Curr Alzheimer Res,2011, 8(1):67-80. [9] Wood TK, Sullivan AM, McDermott KW. Viability of dopaminergic neurones is influenced by serum and astroglial cells in vitro[J]. J Neurocytol, 2003, 32(1):97-103.[10] Zhou Y, Sonobe Y, Akahori T, et al. IL-9 promotes Th17 cell migration into the central nervous system via CC chemokine ligand20 produced by astrocytes[J]. J Immunol, 2011, 186(7): 4415-4421.[11] Kang Z, Altuntas CZ, Gulen MF, et al. Astrocyte-restricted ablation of interleukin-17induced Act1-mediated signaling ameliorates autoimmune encephalomyelitis [J]. Immunity, 2010, 32(3): 414-425. |
|
|
|