# An Edible, Decellularized Plant Derived Cell Carrier for Lab Grown Meat  
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1 Departments of Biomedical Engineering and Biology & Biotechnology, Worcester Polytechnic Institute, Worcester, MA 01609, USA; [EMAIL_ADDRESS] (J.D.J. ); [EMAIL_ADDRESS] (T.D.) [PERSON]  
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[PERSON]  
1 Departments of Biomedical Engineering and Biology & Biotechnology, Worcester Polytechnic Institute, Worcester, MA 01609, USA; [EMAIL_ADDRESS] (J.D.J. ); [EMAIL_ADDRESS] (T.D.) 2  
[PERSON]  
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andrecapitulating the complex native environment of three-dimensional tissues and providing such structured tissues with nutritional and oxygen support beyond their diffusion limits. For these reasons, bioprocess designs specifically concerned with yielding large quantities of cell mass may support a more rapid development of unstructured meat products. One such design, the suspension bioreactor, has been widely employed by the pharmaceutical industry to yield large quantities of cells [5]. Suspension reactors use microcarriers to provide scaffolds for anchorage-dependent cell growth. Such systems are designed to generate fluid flow fields that maximize the number of cells cultured per unit volume while maintaining cell-to-cell homogeneity. A typical bioprocess for culturing mammalian cells in suspension may employ many suspension reactors of increasing size, referred to as seed reactors, within which cells are sequentially expanded to the limits offered by each unit before being inoculated into the larger systems [6]. Stir-tank reactors are especially popular seed-reactors due to their volume capacity, and relatively simple inoculation and harvest procedures; however, their primary criticism, a high media volume to cell quantity ratio, has led researchers to explore the applicability of other bioreactor configurations when operating at scale, such as packed bed and hollow fiber reactors [7]. Many of the target mammalian cell types for lab grown meat are anchorage-dependent and require a stiff substrate upon which they can adhere [8]. Microcarriers are small beads of a wide variety of materials, shapes, sizes, and densities to support the growth of anchorage-dependent cells, such as primary bovine satellite cells, in suspension cultures. Microcarriers have been employed for unique biomedical applications, mainly the production of complex biomolecules such as recombinant proteins; however, there has yet to be a cell carrier designed specifically for lab grown meat [9; 10; 11]. Carriers for use in cellular agriculture must be readily available, affordable, and support cell adhesion and viability during and post inoculation. It is also advantageous to design an edible cell carrier. While cells can be harvested from traditional, inedible carriers, this adds extra steps and associated costs. If carriers are to be integrated into a final unstructured cultured meat product, serving as a bulking agent, a fortifying agent, or a food stabilizer, then they must be both edible and acceptable by consumers. Among plants, broccoli florets have many of the desired characteristics of a cell carrier for cellular agriculture; however, the interactions between native plant cells and seeded mammalian cells remain unknown. Decellularization, a technique used to isolate intact extracellular material from living tissues in their native morphology, has been investigated for applications in tissue engineering [12; 13; 14]. This process involves the exposure of living tissues to a sequence of detergents to remove intracellular components, leaving behind intact extracellular matrix. Decellularization of plants has previously yielded scaffolds that supported adhesion and differentiation of primary bovine satellite cells [15; 16]. Additionally, plant cell walls, the remaining material following plant decellularization process, are primarily composed of cellulose, pectin, and hemicellulose, all of which are considered as nutritionally beneficial in most diets [17].