小胶质细胞
异种移植
移植
生物
流式细胞术
神经科学
干细胞
诱导多能干细胞
细胞分化
细胞生物学
免疫学
医学
胚胎干细胞
炎症
遗传学
基因
外科
作者
Nicola Fattorelli,Anna Martínez-Muriana,Leen Wolfs,Ivana Geric,Bart De Strooper,Renzo Mancuso
出处
期刊:Nature Protocols
[Springer Nature]
日期:2021-01-11
卷期号:16 (2): 1013-1033
被引量:45
标识
DOI:10.1038/s41596-020-00447-4
摘要
Microglia are critically involved in complex neurological disorders with a strong genetic component, such as Alzheimer’s disease, Parkinson’s disease and frontotemporal dementia. Although mouse microglia can recapitulate aspects of human microglia physiology, they do not fully capture the human genetic aspects of disease and do not reproduce all human cell states. Primary cultures of human microglia or microglia derived from human induced pluripotent stem cells (PSCs) are difficult to maintain in brain-relevant cell states in vitro. Here we describe MIGRATE (microglia in vitro generation refined for advanced transplantation experiments, which provides a combined in vitro differentiation and in vivo xenotransplantation protocol to study human microglia in the context of the mouse brain. This article details an accurate, step-by-step workflow that includes in vitro microglia differentiation from human PSCs, transplantation into the mouse brain and quantitative analysis of engraftment. Compared to current differentiation and xenotransplantation protocols, we present an optimized, faster and more efficient approach that yields up to 80% chimerism. To quantitatively assess engraftment efficiency by flow cytometry, access to specialized flow cytometry is required. Alternatively, the percentage of chimerism can be estimated by standard immunohistochemical analysis. The MIGRATE protocol takes ~40 d to complete, from culturing PSCs to engraftment efficiency assessment. Mancuso and De Strooper and colleagues describe MIGRATE (microglia in vitro generation refined for advanced transplantation experiments, a combined in vitro differentiation and in vivo xenotransplantation protocol for studying human microglia transplanted into mouse brain.
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