Laminin enhances the growth of human neural stem cells in defined culture media.

Peter E Hall,Maeve A Caldwell,Justin D Lathia,Charles Ffrench-Constant

doi:10.1186/1471-2202-9-71

Abstract

BackgroundHuman neural stem cells (hNSC) have the potential to provide novel cell-based therapies for neurodegenerative conditions such as multiple sclerosis and Parkinson's disease. In order to realise this goal, protocols need to be developed that allow for large quantities of hNSC to be cultured efficiently. As such, it is important to identify factors which enhance the growth of hNSC. In vivo, stem cells reside in distinct microenvironments or niches that are responsible for the maintenance of stem cell populations. A common feature of niches is the presence of the extracellular matrix molecule, laminin. Therefore, this study investigated the effect of exogenous laminin on hNSC growth.ResultsTo measure hNSC growth, we established culture conditions using B27-supplemented medium that enable neurospheres to grow from human neural cells plated at clonal densities. Limiting dilution assays confirmed that neurospheres were derived from single cells at these densities. Laminin was found to increase hNSC numbers as measured by this neurosphere formation. The effect of laminin was to augment the proliferation/survival of the hNSC, rather than promoting the undifferentiated state. In agreement, apoptosis was reduced in dissociated neurospheres by laminin in an integrin β1-dependent manner.ConclusionThe addition of laminin to the culture medium enhances the growth of hNSC, and may therefore aid their large-scale production.

Highlights

Human neural stem cells have the potential to provide novel cell-based therapies for neurodegenerative conditions such as multiple sclerosis and Parkinson's disease
BMC Neuroscience 2008, 9:71 http://www.biomedcentral.com/1471-2202/9/71 out the brain. They reside within distinct microenvironments or niches such as the ventricular/subventricular zone (VZ/Subventricular zone (SVZ)) of the developing central nervous system (CNS) and the subependymal zone (SEZ) of the adult CNS [1,2]
Human Neural stem cells (NSC) are commonly grown in N2 medium containing 20 ng/ml FGF2 and Epidermal growth factor (EGF), and are passaged by chopping the spheres into segments to be placed in fresh medium

Summary

Introduction

Human neural stem cells (hNSC) have the potential to provide novel cell-based therapies for neurodegenerative conditions such as multiple sclerosis and Parkinson's disease. BMC Neuroscience 2008, 9:71 http://www.biomedcentral.com/1471-2202/9/71 out the brain Instead, they reside within distinct microenvironments or niches such as the ventricular/subventricular zone (VZ/SVZ) of the developing CNS and the subependymal zone (SEZ) of the adult CNS [1,2]. They reside within distinct microenvironments or niches such as the ventricular/subventricular zone (VZ/SVZ) of the developing CNS and the subependymal zone (SEZ) of the adult CNS [1,2] Studies of such niches from different tissues and organisms have led to the identification of common signalling molecules including growth factors, cell-cell signalling molecules, adhesion molecules and extracellular matrix (ECM) molecules, and combinations of these are likely to provide strategies to maintain stem cell populations and so enhance NSC growth in cell culture [3,4]

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