%0 Journal Article %1 ma2004progenitor %A Ma, W. %A Fitzgerald, W. %A Liu, Q.-Y. %A O'Shaughnessy, T.J. %A Maric, D. %A Lin, H.J. %A Alkon, D.L. %A Barker, J.L. %D 2004 %J Experimental Neurology %K 3d methods migration phd stemcell %N 2 %P 276 -- 288 %R http://dx.doi.org/10.1016/j.expneurol.2003.10.016 %T CNS stem and progenitor cell differentiation into functional neuronal circuits in three-dimensional collagen gels %U http://www.sciencedirect.com/science/article/pii/S0014488603005454 %V 190 %X The mammalian central nervous system (CNS) has little capacity for self-repair after injury, and neurons are not capable of proliferating. Therefore, neural tissue engineering that combines neural stem and progenitor cells and biologically derived polymer scaffolds may revolutionize the medical approach to the treatment of damaged \CNS\ tissues. Neural stem and progenitor cells isolated from embryonic rat cortical or subcortical neuroepithelium were dispersed within type I collagen, and the cell–collagen constructs were cultured in serum-free medium containing basic fibroblast growth factor. The collagen-entrapped stem and progenitors actively expanded and efficiently generated neurons, which developed neuronal polarity, neurotransmitters, ion channels/receptors, and excitability. Ca2+ imaging showed that differentiation from BrdU+/TuJ1− to BrdU−/TuJ1+ cells was accompanied by a shift in expression of functional receptors for neurotransmitters from cholinergic and purinergic to predominantly \GABAergic\ and glutamatergic. Spontaneous postsynaptic currents were recorded by patch-clamping from precursor cell-derived neurons and these currents were partially blocked by 10-μM bicuculline, and completely blocked by additional 10 μM of the kainate receptor antagonist CNQX, indicating an appearance of both \GABAergic\ and glutamatergic synaptic activities. Staining with endocytotic marker FM1-43 demonstrated active synaptic vesicle recycling occurring among collagen-entrapped neurons. These results show that neural stem and progenitor cells cultured in 3D collagen gels recapitulate \CNS\ stem cell development; this is the first demonstration of \CNS\ stem and progenitor cell-derived functional synapse and neuronal network formation in a 3D matrix. The proliferative capacity and neuronal differentiating potential of neural progenitors in 3D collagen gels suggest their potential use in attempts to promote neuronal regeneration in vivo.

@article{ma2004progenitor, abstract = {The mammalian central nervous system (CNS) has little capacity for self-repair after injury, and neurons are not capable of proliferating. Therefore, neural tissue engineering that combines neural stem and progenitor cells and biologically derived polymer scaffolds may revolutionize the medical approach to the treatment of damaged \{CNS\} tissues. Neural stem and progenitor cells isolated from embryonic rat cortical or subcortical neuroepithelium were dispersed within type I collagen, and the cell–collagen constructs were cultured in serum-free medium containing basic fibroblast growth factor. The collagen-entrapped stem and progenitors actively expanded and efficiently generated neurons, which developed neuronal polarity, neurotransmitters, ion channels/receptors, and excitability. Ca2+ imaging showed that differentiation from BrdU+/TuJ1− to BrdU−/TuJ1+ cells was accompanied by a shift in expression of functional receptors for neurotransmitters from cholinergic and purinergic to predominantly \{GABAergic\} and glutamatergic. Spontaneous postsynaptic currents were recorded by patch-clamping from precursor cell-derived neurons and these currents were partially blocked by 10-μM bicuculline, and completely blocked by additional 10 μM of the kainate receptor antagonist CNQX, indicating an appearance of both \{GABAergic\} and glutamatergic synaptic activities. Staining with endocytotic marker FM1-43 demonstrated active synaptic vesicle recycling occurring among collagen-entrapped neurons. These results show that neural stem and progenitor cells cultured in 3D collagen gels recapitulate \{CNS\} stem cell development; this is the first demonstration of \{CNS\} stem and progenitor cell-derived functional synapse and neuronal network formation in a 3D matrix. The proliferative capacity and neuronal differentiating potential of neural progenitors in 3D collagen gels suggest their potential use in attempts to promote neuronal regeneration in vivo. }, added-at = {2015-01-06T15:15:05.000+0100}, author = {Ma, W. and Fitzgerald, W. and Liu, Q.-Y. and O'Shaughnessy, T.J. and Maric, D. and Lin, H.J. and Alkon, D.L. and Barker, J.L.}, biburl = {https://www.bibsonomy.org/bibtex/2aedb2b4dfca4e6ab1c1a427f5ae6c062/bkoch}, description = {CNS stem and progenitor cell differentiation into functional neuronal circuits in three-dimensional collagen gels}, doi = {http://dx.doi.org/10.1016/j.expneurol.2003.10.016}, interhash = {1a60c73882f80a164014f8fd1047ceee}, intrahash = {aedb2b4dfca4e6ab1c1a427f5ae6c062}, issn = {0014-4886}, journal = {Experimental Neurology }, keywords = {3d methods migration phd stemcell}, number = 2, pages = {276 -- 288}, timestamp = {2015-05-26T10:46:07.000+0200}, title = {CNS stem and progenitor cell differentiation into functional neuronal circuits in three-dimensional collagen gels }, url = {http://www.sciencedirect.com/science/article/pii/S0014488603005454}, volume = 190, year = 2004 }