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  2. Neural cell isolation from adult macaques for high-throughput analyses and neurosphere cultures

Neural cell isolation from adult macaques for high-throughput analyses and neurosphere cultures

  • Nat Protoc. 2023 Apr 12. doi: 10.1038/s41596-023-00820-z.
Jia-Ru Wei # 1 Dongchang Xiao # 1 Lei Tang # 1 Nana Xu 1 Ruifeng Liu 1 Yuhui Shen 1 Zihui Xu 1 Xuan Sang 1 Jian Ge 1 Mengqing Xiang 1 Sheng Liu 2 3
Affiliations

Affiliations

  • 1 State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China.
  • 2 State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China. liush87@mail.sysu.edu.cn.
  • 3 Guangdong Province Key Laboratory of Brain Function and Disease, Guangzhou, China. liush87@mail.sysu.edu.cn.
  • # Contributed equally.
Abstract

The low number of neural progenitor cells (NPCs) present in the adult and aged primate brains represents a challenge for generating high-yield and viable in vitro cultures of primary brain cells. Here we report a step-by-step approach for the fast and reproducible isolation of high-yield and viable primary brain cells, including mature neurons, immature cells and NPCs, from adult and aged macaques. We describe the anesthesia, transcardial perfusion and brain tissue preparation; the subsequent microdissection of the regions of interest and their enzymatic dissociation, leading to the separation of single cells. The Cell Isolation steps of our protocol can also be used for routine cell culturing, in particular for NPC expansion and differentiation, suitable for studies of hippocampal neurogenesis in the adult macaque brain. The purified primary brain cells are largely free from myelin debris and erythrocytes, paving the way for multiple downstream applications in vitro and in vivo. When combined with single-cell profiling techniques, this approach allows an unbiased and comprehensive mapping of cell states in the adult and aged macaque brain, which is needed to advance our understanding of human cognitive and neurological diseases. The neural Cell Isolation protocol requires 4 h and a team of four to six users with expertize in primary brain Cell Isolation to avoid tissue hypoxia during the time-sensitive steps of the procedure.

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