Shenzhen Key Laboratory for Neuronal Structural Biology, Biomedical Research Institute, Shenzhen Peking University - The Hong Kong University of Science and Technology Medical Center, Guangdong Province, Shenzhen, China.
Shenzhen Key Laboratory for Neuronal Structural Biology, Biomedical Research Institute, Shenzhen Peking University - The Hong Kong University of Science and Technology Medical Center, Guangdong Province, Shenzhen, China; Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay Road, Kowloon, Hong Kong, China.
J Immunol Methods. 2020 Nov;486:112834. doi: 10.1016/j.jim.2020.112834. Epub 2020 Aug 15.
Microglia and astrocytes, the two innate cells in CNS, are thought to protect and remodel of synapses for proper maintenance and plasticity of neuronal circuits. The two types of cells are the major responders by producing and releasing inflammatory mediators. Isolation of microglia and astrocytes from CNS tissue provides a powerful tool to study basic cell biology and examine the effects of in vivo treatments on microglia and astrocytes immunophenotype and function. The widely used approach of enrichment microglia and astrocytes from CNS was MACS (Magnetic activated cell sorting) and FACS (Fluorescence activated cell sorting). Here we described an optimized protocol of enzymatic dissociation generating single cell suspensions from brain tissue. Then the ability of the two methods to isolate microglia and astrocytes from brain dissociated cells was compared. Both MACS and FACS processing could obtain microglia and astrocytes with high viability (>85%). Microglia sorted by MACS comprises a slight myeloid cells contamination but with a little bit higher efficiency than that sorted by FACS. MACS processing was faster than FACS for either single or multiple samples. ACSA2 can be used to isolate astrocytes from both postnatal and adult brain, and is more suitable for purify astrocytes from newborn. FACS could get purer microglia which is helpful for deep sequencing and other related research. ACSA2 is a good marker of astrocytes.
小胶质细胞和星形胶质细胞是中枢神经系统中的两种固有细胞，它们被认为可以保护和重塑突触，以维持神经元回路的适当功能和可塑性。这两种细胞类型通过产生和释放炎症介质成为主要的反应细胞。从小鼠中枢神经系统组织中分离小胶质细胞和星形胶质细胞是研究基础细胞生物学和研究体内治疗对小胶质细胞和星形胶质细胞免疫表型和功能影响的有力工具。MACS（磁激活细胞分选）和 FACS（荧光激活细胞分选）是从小鼠中枢神经系统中富集小胶质细胞和星形胶质细胞的常用方法。在这里，我们描述了一种从脑组织中产生单细胞悬液的优化酶解分离方案。然后比较了这两种方法从小鼠脑分离细胞中分离小胶质细胞和星形胶质细胞的能力。MACS 和 FACS 处理都可以获得高活力（>85%）的小胶质细胞和星形胶质细胞。MACS 分选的小胶质细胞含有少量的髓样细胞污染，但效率略高于 FACS 分选。MACS 处理无论是单个还是多个样本，速度都比 FACS 快。ACSA2 可用于分离出生后和成年期大脑中的星形胶质细胞，并且更适合从新生鼠中纯化星形胶质细胞。FACS 可以获得更纯的小胶质细胞，这有助于深度测序和其他相关研究。ACSA2 是星形胶质细胞的良好标志物。
