Isolation and Characterization of Human Brown Adipocytes
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Isolation and Characterization of Human Brown Adipocytes. / Scheele, Camilla; Henriksen, Tora Ida; Nielsen, Søren.
In: Methods in Molecular Biology, Vol. 2448, 2022, p. 217-234.Research output: Contribution to journal › Journal article › peer-review
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TY - JOUR
T1 - Isolation and Characterization of Human Brown Adipocytes
AU - Scheele, Camilla
AU - Henriksen, Tora Ida
AU - Nielsen, Søren
N1 - Publisher Copyright: © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022
Y1 - 2022
N2 - Brown adipose tissue (BAT) is a thermoregulatory fat with energy-consuming properties. The location and heterogeneity of this tissue makes it complicated to sample before and after interventions in humans, and an in vitro model for mechanistic and molecular studies is therefore of great value. We here describe a protocol for isolation of progenitors from the stromal vascular fraction of BAT biopsies obtained surgically from adult humans. We further present how these cells are differentiated in vitro and finally how they are characterized for thermogenic capacity. Methods for characterization described here include norepinephrine-induced thermogenic gene expression using qPCR; norepinephrine-induced mitochondrial uncoupling using the Seahorse XFe96 Analyzer, and norepinephrine-induced expression of UCP1 using the RNAscope® Technology.
AB - Brown adipose tissue (BAT) is a thermoregulatory fat with energy-consuming properties. The location and heterogeneity of this tissue makes it complicated to sample before and after interventions in humans, and an in vitro model for mechanistic and molecular studies is therefore of great value. We here describe a protocol for isolation of progenitors from the stromal vascular fraction of BAT biopsies obtained surgically from adult humans. We further present how these cells are differentiated in vitro and finally how they are characterized for thermogenic capacity. Methods for characterization described here include norepinephrine-induced thermogenic gene expression using qPCR; norepinephrine-induced mitochondrial uncoupling using the Seahorse XFe96 Analyzer, and norepinephrine-induced expression of UCP1 using the RNAscope® Technology.
KW - Adipogenesis
KW - BAT in vitro model
KW - Brown fat differentiation
KW - Human BAT
KW - Human brown adipocytes
KW - Oxygen consumption rate in human brown adipocytes
KW - UCP1 RNAscope
U2 - 10.1007/978-1-0716-2087-8_14
DO - 10.1007/978-1-0716-2087-8_14
M3 - Journal article
C2 - 35167100
AN - SCOPUS:85124636587
VL - 2448
SP - 217
EP - 234
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -
ID: 299199186