Enhanced neuronal differentiation in a three-dimensional collagen-hyaluronan matrix.
Efficient 3D cell systems for neuronal induction are needed for future use in tissue regeneration. In this study, we have characterized the ability of neural stem/progenitor cells (NS/PC) to survive, proliferate, and differentiate in a collagen type I-hyaluronan scaffold. Embryonic, postnatal, and adult NS/PC were seeded in the present 3D scaffold and cultured in medium containing epidermal growth factor and fibroblast growth factor-2, a condition that stimulates NS/PC proliferation. Progenitor cells from the embryonic brain had the highest proliferation rate, and adult cells the lowest, indicating a difference in mitogenic responsiveness. NS/PC from postnatal stages down-regulated nestin expression more rapidly than both embryonic and adult NS/PC, indicating a faster differentiation process. After 6 days of differentiation in the 3D scaffold, NS/PC from the postnatal brain had generated up to 70% neurons, compared with 14% in 2D. NS/PC from other ages gave rise to approximately the same proportion of neurons in 3D as in 2D (9-26% depending on the source for NS/PC). In the postnatal NS/PC cultures, the majority of betaIII-tubulin-positive cells expressed glutamate, gamma-aminobutyric acid, and synapsin I after 11 days of differentiation, indicating differentiation to mature neurons. Here we report that postnatal NS/PC survive, proliferate, and efficiently form synapsin I-positive neurons in a biocompatible hydrogel.
Brännvall
K
K
Department of Medical Biochemistry and Microbiology, Uppsala University Biomedical Center, Uppsala, Sweden.
Bergman
K
K
Wallenquist
U
U
Svahn
S
S
Bowden
T
T
Hilborn
J
J
Forsberg-Nilsson
K
K
D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D016922:Cellular Senescence; D002540:Cerebral Cortex / Q000166:cytology; D024042:Collagen Type I; D004622:Embryo, Mammalian; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005782:Gels; D006820:Hyaluronic Acid; D051379:Mice; D008822:Mice, Transgenic; D009457:Neuroglia / Q000166:cytology / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D018874:Spheroids, Cellular; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D013696:Temperature
D024042:Collagen Type I; D005782:Gels; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D006820:Hyaluronic Acid