Reconstruction of functional cortical-like tissues from neural stem and progenitor cells.
Neural stem and progenitor cells isolated from embryonic day 13 rat cerebral cortex were immobilized in three-dimensional type I collagen gels, and then the cell-collagen constructs were transferred to rotary wall vessel bioreactors and cultured in serum-free medium containing basic fibroblast growth factor (bFGF) combined with brain-derived neurotrophic factor for up to 10 weeks. Remarkably, the collagen-entrapped cells formed a complex two-layered structure that emulated to a certain extent the cerebral cortex of the embryonic brain in architecture and functionality. The surface layer (layer I) composed primarily of proliferating neural progenitor cells (nestin(+), vimentin(+), and PCNA(+)) predominantly expressed functional neurotransmitter receptors for cholinergic and purinergic agonists while differentiating cells (TuJ1(+) and GFAP(+)) in the deeper layer (layer II) contained differentiated neurons and astrocytes and mainly responded to GABAergic and glutamatergic agonists and to veratridine, which activates voltage-dependent Na(+) channels. An active synaptic vesicle recycling was demonstrated by neuronal networks in the deeper layer using the endocytotic marker FM1-43. Cell polarization forming the characteristic two-layered structure was found to associate with the bFGF and FGF receptor signaling. These engineered functional tissue constructs have a potential use as tissue surrogates for drug screening and detection of environmental toxins, and in neural cell replacement therapy.
Ma
Wu
W
Stem Cell Center, Developmental Biology, American Type Culture Collection (ATCC), Manassas, Virginia 20110, USA. wma@atcc.org
Tavakoli
Tahereh
T
Chen
Silvia
S
Maric
Dragan
D
Liu
Jinny L
JL
O'Shaughnessy
Thomas J
TJ
Barker
Jeffery L
JL
D000818:Animals; D019149:Bioreactors; D016764:Cell Polarity; D024042:Collagen Type I / Q000737:chemistry; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism; D017981:Receptors, Neurotransmitter / Q000378:metabolism; D015222:Sodium Channels / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000295:instrumentation / Q000379:methods
D024042:Collagen Type I; C073804:FM1 43; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D017468:Receptors, Fibroblast Growth Factor; D017981:Receptors, Neurotransmitter; D015222:Sodium Channels