From alpha to beta: biofabricating a pancreatic-like tissue

Abstract

This thesis explores the use of genetic engineering of pancreatic α cells and chaotic bioprinting to develop a pancreatic-like tissue capable of expressing transientnd functional insulin. Since there is not an approach to study or treat type 1 diabetes that merge genetic engineering and bioprinting, we addressed this gap and created this thesis. We successfully develop a novel technique to transfect pancreatic α cells called 3D Lipotransfection, as there is not information regarding this topic on α cells, we took this opportunity and lack of information to contributed to the genetic engineering knowledge. The use of 3D Lipotransfection gave us an improvement in comparison with conventional methods, going from just 32% of efficacy using conventional 2D Lipotransfection, to 81% of efficacy for using 3D Lipotransfection at 72 h post treatment. Insulin-producing α cells demonstrated an outstanding performance to several-dose of glucose stimulation and showed an autocrine regulation to it. The increment of efficacy and cellular confluency of 3D Lipotransfection guided us to a better bioprinted tissue, since the number of cells used is of critical importance in this technique. The utilization of chaotic bioprinting in biological contexts has emerged from a desire to address existing challenges in biofabrication and tissue engineering. Our research demonstrates that chaotic bioprinting and 3D Lipotransfection offers a novel platform for the high-throughput cell culture and genetically modified cells’ maintenance. This work contributes to the initiation of the merge of separated fields and proving better results when this is done.