Long-term three-dimensional neural tissue cultures in functionalized self-assembling peptide hydrogels, matrigel and collagen I

Abstract

Designer peptides with self-assembling properties form nanofibers which are further organized to form a hydrogel consisting of up to 99.5% water. We present here the encapsulation of neural stem cells into peptide nanofiber hydrogel scaffolds. This results in three-dimensional (3-D) neural tissue cultures in which neural stem cells differentiate into progenitor neural cells, neurons, astrocytes and oligodendrocytes when cultured in serum-free medium. Cell survival studies showed that neural cells in peptide hydrogels thrive for at least 5 months. In contrast, neural stem cells encapsulated in Collagen I were poorly differentiated and did not migrate significantly, thus forming clusters. We show that for culture periods of 1-2 weeks, neural stem cells proliferate and differentiate better in Matrigel. However, in long-term studies, the population of cells in Matrigel decreases whereas better cell survival rates are observed in neural tissue cultures in peptide hydrogels. Peptide functionalization with cell adhesion and cell differentiation motifs show superior cell survival and differentiation properties compared to those observed upon culturing neural cells in non-modified peptide hydrogels. These designed 3-D engineered tissue culturing systems have a potential use as tissue surrogates for tissue regeneration. The well-defined chemical and physical properties of the peptide nanofiber hydrogels and the use of serum-free medium allow for more realistic biological studies of neural cells in a biomimetic 3-D environment.