title: Three-dimensional self-organizing neural architectures: a neural stem cells reservoir and a system for neurodevelopmental studies.
abstract: Complex microenvironmental stimuli influence neural cell properties. To study this, we developed a three-dimensional (3-D) neural culture system, composed of different populations including neurons, astrocytes, and neural stem cells (NSCs). In particular, these last-mentioned cells represent a source potentially exploitable to test drugs, to study neurodevelopment and cell-therapies for neuroregenerations. On seeding on matrigel in a medium supplemented with serum and mitogens, cells obtained from human fetal brain tissue formed 3-D self-organizing neural architectures. Immunocytochemical analysis demonstrated the presence of undifferentiated nestin+ and CD133+ cells, surrounded by β-tub-III+ and GFAP+ cells, suggesting the formation of niches containing potential human NSCs (hNSCs). The presence of hNSCs was confirmed by both neurosphere assay and RT-PCR, and their multipotentiality was demonstrated by both immunofluorescent staining and RT-PCR. Flow cytometry analysis revealed that neurosphere forming cells originating from at least two different subsets expressing, respectively, CD133 and CD146 markers were endowed with different proliferative and differentiation potential. Our data implicate that the complexity of environment within niches and aggregates of heterogeneous neural cell subsets may represent an innovative platform for neurobiological and neurodevelopmental investigations and a reservoir for a rapid expansion of hNSCs.
authors: Laboratory of Cellular Neurobiology, Fondazione IRCCS Istituto Neurologico Carlo Besta, Milan, Italy.
authors: Silvia
authors: S
authors: Cristini
authors: Giulio
authors: G
authors: Alessandri
authors: Francesco
authors: F
authors: Acerbi
authors: Emilio
authors: E
authors: Ciusani
authors: Augusto
authors: A
authors: Colombo
authors: Umberto
authors: U
authors: Fascio
authors: Roberto F
authors: RF
authors: Nicosia
authors: Roberto W
authors: RW
authors: Invernizzi
authors: Maurizio
authors: M
authors: Gelati
authors: Eugenio A
authors: EA
authors: Parati
authors: Gloria
authors: G
authors: Invernici
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D001369:Axons / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D001921:Brain / Q000166:cytology / Q000196:embryology; D051929:CD146 Antigen / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005333:Fetus / Q000166:cytology; D005971:Glutamates / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D009420:Nervous System / Q000166:cytology* / Q000254:growth & development*; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D010455:Peptides / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D051929:CD146 Antigen; D005971:Glutamates; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D002118:Calcium