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


RESULTS
To 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 beta1-dependent manner.


CONCLUSIONS
The addition of laminin to the culture medium enhances the growth of hNSC, and may therefore aid their large-scale production.
Hall
Peter E
PE
Department of Pathology, University of Cambridge, Cambridge, UK. peter.hall@keble.ox.ac.uk
Lathia
Justin D
JD
Caldwell
Maeve A
MA
Ffrench-Constant
Charles
C
D017209:Apoptosis / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology; D007797:Laminin / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology*
D003470:Culture Media; D017077:Culture Media, Conditioned; D019012:Integrin beta1; D007797:Laminin; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor