Protocol for isolation of adult mouse ear pinnae-derived primary fibroblasts

• Published in: STAR protocols 2021-06, Vol.2 (2), p.100406-100406, Article 100406
• Main Authors: Bravo, Juan I., Kim, Minhoo, Benayoun, Bérénice A.
• Format: Article
• Language: English
• Subjects: Cell culture; Cell isolation; Protocol
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100406

Researchers need in vitro models that mirror the biology of organisms. Primary fibroblasts play essential roles in wound healing and are present in many tissues. They are widely used in studies of cell cycle control, reprogramming, and aging. Though extraction protocols exist, alternatives that maximize use of available resources are useful. Here, we present our protocol for extracting primary fibroblasts from adult mouse ear pinnae, an often-discarded source of primary cells, which consistently yield large, pure numbers of primary fibroblasts. [Display omitted] •Primary fibroblasts are isolated from adult mouse ear pinnae, an often-unused tissue•Fibroblasts can be isolated from mice of varying sex, age, and genotype•Fibroblast cultures are highly pure, with >90% THY1.2 or PDGFRα staining Researchers need in vitro models that mirror the biology of organisms. Primary fibroblasts play essential roles in wound healing and are present in many tissues. They are widely used in studies of cell cycle control, reprogramming, and aging. Though extraction protocols exist, alternatives that maximize use of available resources are useful. Here, we present our protocol for extracting primary fibroblasts from adult mouse ear pinnae, an often-discarded source of primary cells, which consistently yields large, pure numbers of primary fibroblasts.