Comparative Analysis of Human and Porcine derived Pancreatic dECM using hiPSCs-derived IPCs and LC-MS/MS

Abstract

To recapitulate the microenvironment of the target tissue in the 3D printed tissue construct, the selection of bioink is critical. In previous study, we suggested decellularized extracellular matrix (dECM) bioink as an appropriate material for mimicking the native microenvironment. However, since the source of dECM is generally derived from porcine, it is questionable whether porcine tissue-derived dECM bioink can completely mimic the function of human tissue. In this study, we investigated the differences between human and porcine-derived dECM through a variety of methods to validate that porcine derived dECM provide a suitable microenvironmental cue as for cellular activities, particularly for pancreatic tissue. To evaluate the composition of ECM, we quantified major ECM components before and after decellularization using various biochemical assays. In human and porcine dECM bioinks, the quantity of major components such as collagen and GAGs were observed in similar level. In addition, we conducted comparative analysis of representative components of human and porcine tissue-derived pancreatic dECM using liquid chromatography-mass spectrometry and immunofluorescence staining. The same types of collagen occupied the largest portion of both ECM in common, and other components also appeared in a similar ratio. In addition, we examined the differences of cell-matrix interactions by culturing human-induced pluripotent stem cells (hiPSCs)-derived insulin-producing cells (IPCs) in human and porcine pancreatic dECM (pdECM) bioinks. To assess the effects of the pdECM on cellular function, insulin secretion and gene expression level of IPCs encapsulated in both pdECM bioinks were conducted. These data confirmed that porcine-derived material can also provide beneficial effect under optimized microenvironment condition similar to human tissue. The developed pdECM bioink will be able to broaden the application of in vitro disease models of diabetes and pancreatic cancer and transplantable constructs for in vivo study.