(L-443) Poly-L-Lysine Increases Cell Adhesion to Plant-Based Scaffolds for Tissue Engineering

Introduction:: The conventional animal agriculture industry struggles to meet the needs of worldwide food production. In response to this problem, investigation into alternative agriculture methods such as lab grown meat (cellular agriculture) has become increasingly popular. Decellularized plants have been shown to provide a viable scaffold for cultivating mammalian muscle cells for cellular agriculture when incubated with Fetal Bovine Serum (FBS). FBS is a critical component of eukaryotic cell culture, however, it is expensive and a biproduct of the beef industry, limiting its use. When FBS is removed from cell culture media, eukaryotic muscle cells’ adhesion to decellularized plant scaffolds decreases. We hypothesize coating with positively charged Poly-L-Lysine will increase cell adhesion to decellularized spinach scaffolds when incubated in FBS free media.


Materials and Methods:: QM7 cells (a quail myoblast cell line) in FBS-free-media (DMEM) were seeded onto Poly-L-Lysine coated and uncoated decellularized spinach scaffolds in a 24-well tissue culture plate. The scaffolds were 1⁄2 inch in diameter and each scaffold was seeded with 10,000 QM7 cells. Scaffold samples were fixed with 4% paraformaldehyde at 10, 30, 60, and 120 minutes. The samples were then rinsed with PBS and stained with Hoechst. Images were taken of each scaffold on the BioTek Cytation 1 Cell Imager. Cell count analysis was performed for each sample in ImageJ.

Results, Conclusions, and Discussions:: Results: The cell count analysis showed that on average at 10, 30, 60, and 120 minutes on the uncoated scaffolds, 137 ± 80, 265 ± 238, 254 ± 171, 392 ± 297 cells were attached, respectively. On the Poly-L-Lysine coated scaffolds it was found that on average at 10, 30, 60, and 120 minutes 869 ± 637, 649 ± 413, 1195 ± 1336, 3224 ± 3182 cells were attached, respectively, showing an increase in cell adhesion with Poly-L-Lysine coating and a statistically significant increase in cell adhesion between coated and uncoated groups at the 120-minute time point.



Conclusions: These results indicate that Poly-L-Lysine coating may serve as an effective resource to increase cell adhesion to plant-based scaffolds and improve the efficacy of plant-based scaffolds for the scale-up of cultured meat and tissue engineering.



Discussions: Poly-L-Lysine is cost effective, edible, and commonly used in both cell culture and the food industry. In future studies, we hope to test additional cell types used in cellular agriculture, including bovine satellite cells. Furthermore, we plan to determine the adhesion of cells to decellularized plant scaffolds in a dynamic environment, which will more closely mimic conditions in bioreactors. Given the cost-effectiveness and edibility of Poly-L-Lysine, we suggest coating of decellularized spinach scaffolds can provide an opportunity to reduce costs in the cellular agriculture industry.


Acknowledgements (Optional): : This research is supported by start-up funds to GRG by Boston College. A very special and deserved thank you to Richard Thyden, Luke Perreault, and Glenn Gaudette for their continued guidance and mentorship.

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