Establishment and characterization of primary astrocyte culture from adult mouse brain

星形胶质细胞 生物 胶质纤维酸性蛋白 细胞培养 细胞生物学 神经胶质 胶质细胞源性神经生长因子 基质凝胶 神经营养因子 神经科学 病理 中枢神经系统 免疫学 免疫组织化学 生物化学 医学 受体 遗传学
作者
Xiaoyan Sun,Xin Hu,Dan Wang,Yihang Yuan,Shangyao Qin,Zijian Tan,Yakun Gu,Xu Huang,Cheng He,Zhida Su
出处
期刊:Brain Research Bulletin [Elsevier BV]
卷期号:132: 10-19 被引量:28
标识
DOI:10.1016/j.brainresbull.2017.05.002
摘要

As a major class of glial cells, astrocytes have been indicated to play multi-roles in physiological and pathological brain. Astrocyte cultures derived from postnatal mouse brains have been extensively used to characterize their biological properties. However, the inability to culture adult mouse primary astrocytes has long stymied studies of function in adult brain. Here, we developed a protocol to successfully establish highly enriched astrocyte cultures from the brains of adult mouse. Cortical tissues were collected to prepare cell suspension by enzymatic digestion and mechanical dissociation, and then plated onto vessels pre-coated with gelatin and matrigel and cultured in DMEM medium containing 20% fetal bovine serum (FBS). Forskolin (FSK) and glial-derived neurotrophic factor (GDNF) were use to promote astrocyte proliferation and survival respectively. These adult astrocyte cultures were identified by immunocytochemical, immunobloting and PCR analysis. Furthermore, biological and functional analysis indicated that they possess the biochemical and physiological properties of astrocytes, suggestive of a useful cell model for astroglial studies in adult brain.
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