Protocol to maintain single functional mouse hematopoietic stem cells in vitro without cell division

Belmonte, Miriam; Kent, David G.

doi:10.1016/j.xpro.2021.100927

STAR Protocols

2021

DOI: 10.1016/j.xpro.2021.100927

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Protocol to maintain single functional mouse hematopoietic stem cells in vitro without cell division

Miriam Belmonte

David G. Kent

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Cited by 2 publications

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“…A deep understanding of the HSC metabolism will provide novel insights into the regulation of hematopoietic homeostasis and leukemic transformation and aid in improving clinical applications such as ex vivo expansion for transplantation purposes and therapeutic treatment schemes. Depending on the surface markers used for identification, between 1000 to 5000 HSCs can be isolated from a single mouse. − …”

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confidence: 99%

LC-MS-Based Targeted Metabolomics for FACS-Purified Rare Cells

et al. 2023

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Metabolism plays a fundamental role in regulating cellular functions and fate decisions. Liquid chromatography-mass spectrometry (LC-MS)-based targeted metabolomic approaches provide high-resolution insights into the metabolic state of a cell. However, the typical sample size is in the order of 10 5 −10 7 cells and thus not compatible with rare cell populations, especially in the case of a prior flow cytometry-based purification step. Here, we present a comprehensively optimized protocol for targeted metabolomics on rare cell types, such as hematopoietic stem cells and mast cells. Only 5000 cells per sample are required to detect up to 80 metabolites above background. The use of regularflow liquid chromatography allows for robust data acquisition, and the omission of drying or chemical derivatization avoids potential sources of error. Cell-type-specific differences are preserved while the addition of internal standards, generation of relevant background control samples, and targeted metabolite with quantifiers and qualifiers ensure high data quality. This protocol could help numerous studies to gain thorough insights into cellular metabolic profiles and simultaneously reduce the number of laboratory animals and the time-consuming and costly experiments associated with rare cell-type purification.

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LC-MS-Based Targeted Metabolomics for FACS-Purified Rare Cells

et al. 2023

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IL‐11—An aging‐related cytokine with opportunities for regulating hematopoiesis

Kent

2024

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Protocol to maintain single functional mouse hematopoietic stem cells in vitro without cell division

Cited by 2 publications

References 12 publications

LC-MS-Based Targeted Metabolomics for FACS-Purified Rare Cells

LC-MS-Based Targeted Metabolomics for FACS-Purified Rare Cells

IL‐11—An aging‐related cytokine with opportunities for regulating hematopoiesis

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