# An Edible, Decellularized Plant Derived Cell Carrier for Lab Grown Meat
## 4 Discussion
The relatively high eccentricity of the decellularized florets, when compared to traditional, spherical microcarriers might be advantageous, as shear stresses have been shown to promote alignment of mammalian cells, a necessary characteristic of functional muscle tissue [35]. The applicability of decellularized plant-derived cell carriers, in consideration of a scaled bioprocess, was studied. The decellularization process used in this paper reflected a suspension-style method previously reported; however, a few changes were made [16]. First, Triton X-100, which is not generally regarded as safe in food products was replaced with Polysorbate-20 (Tween), an emulsifier that has been approved as safe by the Food and Drug Administration [36]. Secondly, in the interest of reducing the water use and time to yield decellularized plants, SDS, Polysorbate-20, and bleach were combined into one decellularization solution. This decreased the processing time from 7 days to 2 days and water use by a factor of 3.5. Further optimization of this decellularization process is encouraged for both broccoli and other widely available plant materials. The use of plant waste as cell carriers, such as wheat middlings, a byproduct of wheat processing, or aquatic plant waste, which is typically displaced via dredging or beach combing, may further support sustainability efforts. The development of edible, nutritionally beneficial, scaffolds as presented in this study might further simplify a scaled cultured meat bioprocess. Synthetic microcarriers are inedible and would require a potentially expensive harvest procedure to ensure no inedile materials were incorporated into the final product. This harvest procedure may require digestive enzymes, such as animal derived trypsin. The purification of trypsin, which is dependent on secondary bioprocesses, will come with an associated cost, estimated at approximately 98 USD/liter [37]. Additionally, trypsin harvesting from small scale microcarrier suspension cultures is reported to vary between 70-95%, suggesting that this step may waste cell biomass [38]. Figure 7: Comparison of the density of decellularized florets to microcarriers currently used in practice for biological applications. Density measurements for decellularized florets were performed by our lab. Density measurements for Cytodex 1, Cytodex 3, Cytopore 1, and Cultsipher S, are previously reported [30; 31; 32; 33]. Such processes will drive up the cost of manufacturing for a final product. While there are commercial microcarriers derived from edible materials, they were not designed in consideration of their nutritional composition. For example, Cytodex microcarriers are composed of dextran which is believed to convert to glucose upon ingestion, and may ultimately be a health detriment to diabetic consumers [39]. The cell carriers proposed in this paper, being composed primarily of the insoluble dietary fiber cellulose, have well documented positive effect on health [17]. Additionally, research into biofortification has yielded procedures to improve the nutritional characteristics of fresh plants. If these fortifications can be maintained through the decellularization process, the nutritional profile of a final product can be further improved [40].