Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates

从非人灵长类动物的胎儿骨骼和肝脏中分离单核细胞,用于代谢、功能和免疫表型分析。

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作者:Michael J Nash ,Evgenia Dobrinskikh ,Dong Wang ,Eric M Pietras ,Rachel C Janssen ,Jacob E Friedman ,Stephanie R Wesolowski

Abstract

Studying fetal hematopoiesis is challenging as hematopoiesis transitions from the liver to bone marrow. Obtaining human samples is not possible, and small animal models may not provide sufficient biological material. Here, we present a protocol for isolating hematopoietic cells from the nonhuman primate fetal liver and bone. We describe steps for using cells from the same fetus for fluorescence lifetime imaging microscopy to measure metabolism, assessing cellular function, and flow cytometry for immunophenotyping at the single-cell level. For complete details on the use and execution of this protocol, please refer to Nash et al. (2023).1.

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