FibroPrep protocol for rapid enzyme-free isolation of primary dermal fibroblasts from adult mouse skin for biological assays

Isolating primary dermal fibroblasts is often costly, time-consuming, has high contamination risk, and is inefficient, yielding low cell numbers and requiring long culture periods. Here, we present a non-enzymatic isolation protocol from mouse skin that overcomes these limitations and consistently provides passage 1 (P1) primary fibroblasts with every tissue subculture. We describe steps for isolating primary fibroblasts from adult mice dermis by placing a precise small dermal tissue layer, which adhered after 24 h, with migrating fibroblasts observed at 48 h post isolation.