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Tissue Eng Part C Methods. 2015 Mar;21(3):314-21. doi: 10.1089/ten.TEC.2014.0118. Epub 2014 Dec 17.

Live fibroblast harvest reveals surface marker shift in vitro.

Author information

1
1 Hagey Laboratory for Pediatric Regenerative Medicine, Department of Surgery, Plastic and Reconstructive Surgery, Stanford University School of Medicine , Stanford, California.

Abstract

Current methods for the isolation of fibroblasts require extended ex vivo manipulation in cell culture. As a consequence, prior studies investigating fibroblast biology may fail to adequately represent cellular phenotypes in vivo. To overcome this problem, we describe a detailed protocol for the isolation of fibroblasts from the dorsal dermis of adult mice that bypasses the need for cell culture, thereby preserving the physiological, transcriptional, and proteomic profiles of each cell. Using the described protocol we characterized the transcriptional programs and the surface expression of 176 CD markers in cultured versus uncultured fibroblasts. The differential expression patterns we observed highlight the importance of a live harvest for investigations of fibroblast biology.

PMID:
25275778
PMCID:
PMC4346232
DOI:
10.1089/ten.TEC.2014.0118
[Indexed for MEDLINE]
Free PMC Article

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