This study evaluates 32 isolation conditions using four enzyme mixtures (Collagenase type I, Collagenase type I + Trypsin, Liberase™, and Collagenase type IV) at varying concentrations and incubation times to isolate bovine mesenchymal stromal cells (MSCs) from adipose tissue. Liberase™ at 0.1% for 3 hours yielded the highest cell count with a low population doubling time. MSC identity was confirmed, and myogenic differentiation was achieved using 5-aza-2'-deoxycytidine and galectin-1, increasing myogenic regulatory factor mRNA levels. For cultured meat production using bovine AT-MSCs, 0.1% Liberase™ for 3 hours is recommended.

Emma Heyman, Bert Devriendt, Elly De Vlieghere, Klara Goethals, Mario Van Poucke, Luc Peelman, Catharina De Schauwer