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的两个先天细胞，被认为可以保护和重塑突触，从而适当维持神经元回路的可塑性。两种类型的细胞是通过产生和释放炎性介质的主要反应物。从中枢神经系统组织中分离出小胶质细胞和星形胶质细胞，为研究基础细胞生物学和检查体内治疗对小胶质细胞和星形胶质细胞免疫表型和功能的影响提供了强大的工具。从中枢神经系统富集小胶质细胞和星形胶质细胞的广泛使用的方法是MACS（磁激活细胞分选）和FACS（荧光激活细胞分选）。在这里，我们描述了从脑组织产生单细胞悬液的酶解的优化方案。然后比较了两种方法从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处理均可获得具有高生存力（> 85％）的小胶质细胞和星形胶质细胞。通过MACS分选的小胶质细胞含有轻微的髓样细胞污染，但效率比通过FACS分选的小胶质细胞高一点。对于单个或多个样本，MACS处理都比FACS更快。ACSA2可用于从出生后和成年大脑中分离星形胶质细胞，并且更适合从新生儿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记。通过MACS分选的小胶质细胞含有轻微的髓样细胞污染，但效率比通过FACS分选的小胶质细胞高一点。对于单个或多个样本，MACS处理都比FACS更快。ACSA2可用于从出生后和成年大脑中分离星形胶质细胞，并且更适合从新生儿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记。通过MACS分选的小胶质细胞含有轻微的髓样细胞污染，但效率比通过FACS分选的小胶质细胞高一点。对于单个或多个样本，MACS处理都比FACS更快。ACSA2可用于从出生后和成年大脑中分离星形胶质细胞，并且更适合从新生儿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记。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记。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记。