Facile formation of zein microbeads integrated with a plant protein-based bioink for cultivated meat texturization

The cultivated meat (CM) field is advancing rapidly, yet scalable expansion of muscle and adipose precursor cells and their subsequent assembly into CM products with desirable organoleptic and nutritional properties remain challenging. Edible microbeads (MBs) represent promising microcarriers for large-scale cell expansion in bioreactors, as they can be directly incorporated into final products. However, most existing MBs are derived from animal proteins, such as gelatin, which conflicts with the goal of producing sustainable and affordable CM products. Here, we report the fabrication of pure plant-protein MBs from zein, a corn-derived prolamin, with tunable sizes of 100–500 μm. Using a fully food-grade, scalable emulsion–evaporation method, we produced kilogram-level zein MBs that supported cell culture in spinner-flask bioreactors without additional cross-linking or surface modification. These MBs enabled robust expansion of porcine skeletal muscle satellite cells (PSCs) and adipose-derived mesenchymal stem cells (ADSCs), while also serving as functional components in the formulation of novel mung bean protein/alginate (MA) edible inks. By encapsulating cell-laden zein MBs within MA ink, we employed a dual-nozzle extrusion bioprinter to fabricate centimeter-scale marbled CM constructs that recapitulate the appearance, texture, nutritional, and flavor profile of pork belly, as confirmed by instrumental analyses. This work establishes a commercially viable plant protein-based platform that integrates zein MBs and mung bean protein ink with dynamic bioreactors and 3D bioprinting, enabling scalable cell expansion and the fabrication of customizable CM products with defined textures. • Fabrication of Zein MBs (100–500 μm) via a scalable emulsion–evaporation method • Zein MBs act as edible microcarriers for muscle and adipose stem cell expansion • Zein MB–loaded mung bean protein/alginate inks show favorable printability • Cell-laden inks enable texturization of marbled cultivated meat via 3D bioprinting