title: Live fibroblast harvest reveals surface marker shift in vitro.
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.
authors.affiliation: 1 Hagey Laboratory for Pediatric Regenerative Medicine, Department of Surgery, Plastic and Reconstructive Surgery, Stanford University School of Medicine , Stanford, California.
authors.forename: Graham G
authors.initials: GG
authors.lastname: Walmsley
authors.forename: Yuval
authors.initials: Y
authors.lastname: Rinkevich
authors.forename: Michael S
authors.initials: MS
authors.lastname: Hu
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Montoro
authors.forename: David D
authors.initials: DD
authors.lastname: Lo
authors.forename: Adrian
authors.initials: A
authors.lastname: McArdle
authors.forename: Zeshaan N
authors.initials: ZN
authors.lastname: Maan
authors.forename: Shane D
authors.initials: SD
authors.lastname: Morrison
authors.forename: Dominik
authors.initials: D
authors.lastname: Duscher
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Whittam
authors.forename: Victor W
authors.initials: VW
authors.lastname: Wong
authors.forename: Irving L
authors.initials: IL
authors.lastname: Weissman
authors.forename: Geoffrey C
authors.initials: GC
authors.lastname: Gurtner
authors.forename: Michael T
authors.initials: MT
authors.lastname: Longaker
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology*; D005434:Flow Cytometry; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D046509:Tissue Culture Techniques; D014158:Transcription, Genetic
chemical_list: D015415:Biomarkers