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Flow Cytometry Has a Significant Impact on the Cellular Metabolome.
Binek, Aleksandra; Rojo, David; Godzien, Joanna; Rupérez, Francisco Javier; Nuñez, Vanessa; Jorge, Inmaculada; Ricote, Mercedes; Vázquez, Jesús; Barbas, Coral.
Afiliação
  • Binek A; Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III , Madrid 28029 , Spain 1.
  • Rojo D; Centro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia , Universidad CEU San Pablo , Campus Montepríncipe , Madrid 28668 , Spain.
  • Godzien J; Centro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia , Universidad CEU San Pablo , Campus Montepríncipe , Madrid 28668 , Spain.
  • Rupérez FJ; Centro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia , Universidad CEU San Pablo , Campus Montepríncipe , Madrid 28668 , Spain.
  • Nuñez V; Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III , Madrid 28029 , Spain 1.
  • Jorge I; Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III , Madrid 28029 , Spain 1.
  • Ricote M; CIBER de Enfermedades Cardiovasculares (CIBER CV), Madrid 28029 , Spain.
  • Vázquez J; Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III , Madrid 28029 , Spain 1.
  • Barbas C; Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III , Madrid 28029 , Spain 1.
J Proteome Res ; 18(1): 169-181, 2019 01 04.
Article em En | MEDLINE | ID: mdl-30362351
ABSTRACT
The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyze the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites related to the plasma membrane, activating a mechanosensory signaling cascade causing inflammation-like stress. The procedure also triggered alterations related to energy consumption and cell damage. The observed changes mostly derive from the physical impact on cells during their passage through the instrument. These findings provide evidence of FACS-induced biochemical changes, which should be taken into account in the design of robust metabolic assays of cells separated by flow cytometry.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Separação Celular / Metaboloma / Citometria de Fluxo Limite: Animals Idioma: En Ano de publicação: 2019 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Separação Celular / Metaboloma / Citometria de Fluxo Limite: Animals Idioma: En Ano de publicação: 2019 Tipo de documento: Article