Evaluation of food-grade antifoams for biocompatibility, differentiation, and bioprocess performance in porcine mesenchymal stem cells cultivation for cellular agriculture

Background Cultivated meat production requires scalable livestock stem cells expansion, but foaming during cultivation in bioreactors remains a critical challenge. Protein-rich media, high agitation, and gas sparging promote foam formation, reducing oxygen transfer, altering hydrodynamics, and threatening sterility. Chemical-grade antifoams like Poloxamer 188 are widely used in mammalian cell culture, but their cost and lack of food-grade status limit suitability for cellular agriculture. Objective We evaluated food-grade antifoams in the culture of primary porcine adipose-derived mesenchymal stem cells (pAD-MSCs). Methods We assessed cytotoxicity, plastic adherence, aggregate formation, bioreactor foam suppression, and adipogenic differentiation across concentrations. Results Several candidates showed no toxicity and even enhanced viable cell numbers, most silicone/ polydimethylsiloxane-based (PDMS) agents showed time- and dose-dependent toxicity. One formulation enhanced adipogenic differentiation compared to the control. Antifoams reduced spheroid size without impairing aggregation and decreased 2D adherence. All non-toxic candidates effectively suppressed foam at working concentrations. Conclusions Food-grade antifoams robust foam control in pAD-MSCs workflows without major toxicity and potential benefits for differentiation. Because EU Novel Food authorization requires ingredient-level safety and process transparency, edible and cost-effective antifoams may be advantageous candidates for future regulatory and process development.