Journal
DEVELOPMENTAL BRAIN RESEARCH
Volume 153, Issue 2, Pages 163-173Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.devbrainres.2004.08.010
Keywords
cell aggregate; neural stem cell; differentiation; 3D collagen; rotating wall vessel
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Neural stem cells and neural progenitors (NSCs/NPs) are capable of self-renewal and can give rise to both neurons and glia. Such cells have been isolated from the embryonic brain and immobilized in three dimensional collagen gels. The collagen-entrapped NSCs/NPs recapitulate CNS stem cell development and form functional synapses and neuronal circuits. However, the cell-collagen constructs from static conditions contain hypoxic, necrotic cores and the cells are short-lived. In the present study, NSCs/NPs isolated from embryonic day 13 rat cortical neuroepithelium are immobilized in type 1 collagen gels and cultured in NASA-designed rotating wall vessel (RWV) bioreactors for up to 9 weeks. Initially, during the first 2 weeks of culture, a lag phase of cellular growth and differentiation is observed in the RWV bioreactors. Accelerated growth and differentiation, with the cells beginning to form large aggregates (similar to1 mm in diameter) without death cores, begins during the third week. The collagen-entrapped NSCs/NPs cultured in RWV show active neuronal generation followed by astrocyte production. After 6 weeks in rotary culture, the cell-collagen constructs contain over 10 fold greater nestin(+) and GFAP(+) cells and two-fold more TuJ1 gene expression than those found in static cultures. In addition, TuJ1(+) neurons in RWV culture give rise to extensive neurite outgrowth and considerably more synapsin 1(+) pre-synaptic puncta surrounding MAP2(+) cell bodies and dendrites. These results strongly suggest that the cell-collagen-bioreactor culture system supports long-term NSC/NP growth and differentiation, and RWV bioreactors can be useful in generating neural tissue like constructs, which may have the potential for cell replacement therapy. (C) 2004 Elsevier B.V. All rights reserved.
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