HEPATOCYTES CULTURE IN SELF-ASSEMBLED PEPTIDE HYDROGELS

Abstract

The development and research of new alternatives to drugs testing in animals is a priority as an ethical matter, several researchers have been working in developing new ways for toxicity testing in 3D in vitro models. The self-assembling process of short peptide sequences has been demonstrated to form nanofibrous hydrogels, this may provide an acceptable alternative, since it is known that the majority of cells from the human body are surrounded by a nanofibrous extracellular matrix (ECM). Peptide hydrogels have been described to work by mimicking the extracellular matrix and provide a suitable 3D environment for in vitro cell culture (Gough, et al., 2011). In this research, five different novel self-assembling peptide hydrogels were investigated to find the most suitable ECM substitute and whether the hydrogels could support the growth and maintain the function of liver hepatocytes; the viability of cells was tested using a live/dead assay at three different time points (1, 3 and 7 days); hepatocytes were cultured on the surface of preformed gels (2D environment) and also encapsulated within the gel structure (3D environment), the latter case more closely mimicking the ECM. After a qualitative comparison of the viability and cytotoxicity between the 5 different peptide sequences the selected hydrogel was characterised as a 3D environment using 3 different cell densities (5 x 105 cells mL-1, 1 x 106 cells mL-1 and 2 x 106 cells mL-1); Since the focus was using hepatocytes, the ability of the hydrogels to provide a suitable and supporting environment was assessed by the measurement of albumin and urea production which are major indicators of whether hepatocytes maintained their phenotype within the gels.

Date of Award	1 Aug 2016
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Julie Gough (Supervisor) & Aline Saiani (Supervisor)