FibroPrep protocol for rapid enzyme-free isolation of primary dermal fibroblasts from adult mouse skin for biological assays
Year: 2025
Type: article
Abstract: 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. • Steps for enzyme-free isolation of murine dermal fibroblasts • Procedures for subculturing and maintaining a continuous passage 1 fibroblast culture • Details on assessing the purity and integrity of NEDI-isolated primary dermal fibroblasts Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.
Source: STAR Protocols
Institution: Indian Institute of Technology Kharagpur
Language: English
Topic: Wound Healing and Treatments
Subfield: Rehabilitation
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W7116765472