Optimization of Matrigel-based culture for expansion of neural stem cells

Neural stem cells (NSCs) are capable of self-renewal and can differentiate into neurons and two types of glial cells. NSCs have great potential for basic studies of neurogenesis and neurodegenerative diseases, and for therapeutic transplantation applications. NSCs can be derived from the fetal or adult brain. In vitro expansion of NSCs may help to elucidate their properties involved in neurogenesis and is crucial to obtain on-demand the large numbers of these cells needed for clinical transplantation trials. Laminin is an extracellular matrix molecule commonly used to culture NSCs as an attached monolayer. However, laminin is costly if NSCs need to be cultured in large quantities. Matrigel is a soluble basement membrane biomaterial, which has been widely used for feeder-free cultures of human embryonic stem cells. In the present study, we evaluated the ability of Matrigel to support the attachment and long-term proliferation of NSCs. We investigated the growth and adherence of NSCs derived from the fetal mouse brain on Matrigel at various concentrations (0.01–1%). We found that 0.02% Matrigel is sufficient for NSCs to be expanded in vitro in long-term culture. The NSCs cultured on Matrigel-coated plates showed typical cellular and molecular characteristics and had multipotency to be differentiated into neurons and glial cells. The Matrigel-based monolayer culture system established in this study provides a cost-effective approach to maintain NSCs in vitro in long-term cultures.