Conditioning three-dimensional biomimetic scaffolds for the cultivation of transplantable beta cells

Aims: Islet transplantation holds vast potential as a treatment for Type 1 diabetes and provides recipients with short-term insulin independence. A major limitation of this treatment is the lack of donor beta cells available for transplantation. Current two-dimensional cell culture methods have generated cell populations which possess a functioning beta cell phenotype. However, these cells are not suitable for clinical transplantation. Three-dimensional culture methods that more accurately mimic the complexity of the in vivo pancreatic environment may offer a new platform on which an alternative source of transplantable cell populations can be differentiated and cultured. Methods: Populations of 1 9 106 Min-6 were seeded onto three-dimensional porous BioVyonTM scaffolds and cultured for up to 15 days. The scaffolds were incubated with fetal bovine serum, collagen IV and laminin for 2h prior to seeding. Cell populations were exposed to a glucose challenge at 3, 9 and 15 days prior to quantification using an LDH assay. Populations were stained using Calcein AM and Hoescht and propidium iodide to ascertain viability. Results: Min-6 populations were observed to proliferate and remained viable over 15 days within the BioVyonTM environment. A significant increase in the quantities of insulin secreted to fluctuating glucose concentrations was observed over time (p < 0.001). Conclusion: Modification of the BioVyonTM surface with proteins found in the peri-insular basement membrane was successful in harbouring and sustaining large populations of the Min-6 cell line. The protected beta cell phenotype observed in the Min-6 populations suggests that BioVyonTM could hold potential as a stem cell culture and differentiation platform.