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从胎儿骨骼和肝脏中分离单核细胞,用于非人类灵长类动物的代谢、功能和免疫表型分析。

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

机构信息

Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.

Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.

出版信息

STAR Protoc. 2024 Mar 15;5(1):102849. doi: 10.1016/j.xpro.2024.102849. Epub 2024 Feb 6.

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)..

摘要

研究胎儿造血是具有挑战性的,因为造血从肝脏向骨髓转变。获得人体样本是不可能的,而小动物模型可能无法提供足够的生物材料。在这里,我们提出了一种从非人灵长类动物胎儿肝脏和骨骼中分离造血细胞的方案。我们描述了使用同一胎儿的细胞进行荧光寿命成像显微镜测量代谢、评估细胞功能以及单细胞水平的免疫表型流式细胞术的步骤。有关该方案使用和执行的完整详细信息,请参阅 Nash 等人。(2023 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2286/10859273/1eda6b5de6b3/fx1.jpg

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