Functionalization of Chitosan Based Microparticles for In Vitro 3D Culture of Human Liver Cells

Abstract

Previous work involving 3D culture of human liver cells with fluorinated chitosan based microparticles has shown that their incorporation provides needed structural cues that the culture of cells alone does not provide, such as increased gas transport. However, using bare microparticles to grow large 3D cellular structures is not practical as they tend to collapse before any meaningful research can be done on them. It is proposed that this is because of the lack of extracellular matrix (ECM)components within pure cell culture in vitro, which in their absence cannot adequately facilitate biochemical communication and adhesion between cells. It is possible to utilize protein linkage techniques, namely carbodiimide chemistry, to immobilize proteins derived from the liver cellular environment to the surface of these microparticles and grow stable 3D cell aggregates known as spheroids. I hypothesized that the surface functionalization of fluorinated methacrylamide chitosan (MACF) microparticles with liver ECM adhesive ligands and their culture with liver cells will increase formation and biological function of spheroids. It was found that through the modification of MACF microparticles with native extracellular matrix proteins, particularly variations of laminin native to the in vivo liver environment, the overall functionality of cells increased over a 5 day culture time, as shown by higher levels of both albumin and urea secretion when compared to controls (p\u3c0.05)

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