Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions

Summary: Here, we present a protocol for differentiating 3T3-L1 preadipocytes and stromal vascular fraction (SVF)-derived preadipocytes from mice into mature adipocytes, followed by the isolation of crude mitochondrial fractions. This cost-effective and reproducible protocol is optimized for small-p...

Full description

Saved in:
Bibliographic Details
Main Authors: Churaibhon Wisessaowapak, Jeongmin Lee, Hyeonhui Kim, Seunghwan Son, Xue Feng, Lina Chang, Annie Hoang, Hetty Chen, Sarah Bedsted, Alan R. Saltiel
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725004514
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849225298719014912
author Churaibhon Wisessaowapak
Jeongmin Lee
Hyeonhui Kim
Seunghwan Son
Xue Feng
Lina Chang
Annie Hoang
Hetty Chen
Sarah Bedsted
Alan R. Saltiel
author_facet Churaibhon Wisessaowapak
Jeongmin Lee
Hyeonhui Kim
Seunghwan Son
Xue Feng
Lina Chang
Annie Hoang
Hetty Chen
Sarah Bedsted
Alan R. Saltiel
author_sort Churaibhon Wisessaowapak
collection DOAJ
description Summary: Here, we present a protocol for differentiating 3T3-L1 preadipocytes and stromal vascular fraction (SVF)-derived preadipocytes from mice into mature adipocytes, followed by the isolation of crude mitochondrial fractions. This cost-effective and reproducible protocol is optimized for small-plate formats, compatible with standard reagents, and suitable for metabolic studies such as insulin resistance and mitochondrial function. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
format Article
id doaj-art-faaec5f58d5e46fbbd3cafd55cedf191
institution Kabale University
issn 2666-1667
language English
publishDate 2025-09-01
publisher Elsevier
record_format Article
series STAR Protocols
spelling doaj-art-faaec5f58d5e46fbbd3cafd55cedf1912025-08-25T04:14:48ZengElsevierSTAR Protocols2666-16672025-09-016310404510.1016/j.xpro.2025.104045Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractionsChuraibhon Wisessaowapak0Jeongmin Lee1Hyeonhui Kim2Seunghwan Son3Xue Feng4Lina Chang5Annie Hoang6Hetty Chen7Sarah Bedsted8Alan R. Saltiel9Division of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USA; Corresponding authorDivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USADivision of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, San Diego, CA, USA; Department of Pharmacology, University of California, San Diego, San Diego, CA, USA; Corresponding authorSummary: Here, we present a protocol for differentiating 3T3-L1 preadipocytes and stromal vascular fraction (SVF)-derived preadipocytes from mice into mature adipocytes, followed by the isolation of crude mitochondrial fractions. This cost-effective and reproducible protocol is optimized for small-plate formats, compatible with standard reagents, and suitable for metabolic studies such as insulin resistance and mitochondrial function. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725004514Cell cultureMetabolismMolecular Biology
spellingShingle Churaibhon Wisessaowapak
Jeongmin Lee
Hyeonhui Kim
Seunghwan Son
Xue Feng
Lina Chang
Annie Hoang
Hetty Chen
Sarah Bedsted
Alan R. Saltiel
Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions
STAR Protocols
Cell culture
Metabolism
Molecular Biology
title Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions
title_full Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions
title_fullStr Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions
title_full_unstemmed Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions
title_short Protocol for differentiating murine 3T3-L1 and SVF-derived preadipocytes and isolating crude mitochondrial fractions
title_sort protocol for differentiating murine 3t3 l1 and svf derived preadipocytes and isolating crude mitochondrial fractions
topic Cell culture
Metabolism
Molecular Biology
url http://www.sciencedirect.com/science/article/pii/S2666166725004514
work_keys_str_mv AT churaibhonwisessaowapak protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT jeongminlee protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT hyeonhuikim protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT seunghwanson protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT xuefeng protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT linachang protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT anniehoang protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT hettychen protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT sarahbedsted protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions
AT alanrsaltiel protocolfordifferentiatingmurine3t3l1andsvfderivedpreadipocytesandisolatingcrudemitochondrialfractions