Ex Vivo Expansion and Cryopreservation of Goat Satellite Cells

The aim of this study was to develop an improvised protocol for ex vivo expansion of caprine satellite cells (SCs) using growth factors and their cryopreservation. The SCs were isolated from muscle tissue using collagenase type-I for enzymatic digestion. In vitro culture of SCs was done using Dulbecco's modified Eagle medium (DMEM) supplemented with 20% fetal bovine serum (FBS) and 15% horse serum. Immunofluorescence and PCR confirmed positive expression of PAX7 and MyoD, and negative expression of MyHC. Supplementation with 10 ng/ml basic fibroblast growth factor (bFGF) + 100 ng/ml insulin-like growth factor-1 (IGF-1) in the basal culture medium (DMEM + 2% FBS) showed similar results as control (routine culture medium). Subsequently, SCs were successfully cryopreserved using 10% dimethyl sulfoxide and 40% FBS with post-thaw viability of 74.72%. We have successfully established a protocol for the isolation, culture, and cryopreservation of goat SCs. Caprine SCs can be successfully expanded in vitro using growth factors (10 ng/ml bFGF and 100 ng/ml IGF-1) with minimum supplemental FBS in basal culture medium. Additionally, investigations could assess the viability and integrity of cryopreserved goat SCs post-thaw, ensuring that their proliferative capabilities remain intact. © 2025 Wiley Periodicals LLC. Basic Protocol: Isolation, culture, cost-effective expansion, and cryopreservation of caprine satellite cells.