Alignment of glial cells stimulates directional neurite growth of CNS neurons in vitro.
Olfactory ensheathing cells (OECs) together with olfactory nerve fibroblasts (ONFs) and neonatal astrocytes are potent stimulators of neurite growth in adulthood and during development, respectively. Since it is known that alignment of glial cells is important for the correct outgrowth of axon tracts, it was hypothesized that the alignment of glial cells stimulates directional and enhanced neurite outgrowth. Adult OEC/ONF and neonatal astrocytes were cultured either on biodegradable poly(d,l)-lactide matrices or in Petri dishes for 4 days. Thereafter neonatal cerebral cortical neurons were added. After a 2-days coculture period the cultures were fixed and processed for a combined MAP-2 and phosphorylated neurofilament (RT97) staining. The neurite growth (neurite elongation and neurite formation) and the neurite direction were assessed. We show that (1). OEC/ONF cultures are more potent in stimulating the length of the longest neurite of cocultured neurons, (2). alignment of glial is achieved in vitro on our biomatrices, (3). aligned glial/biomatrix complexes do not enhance neurite growth, and (4). aligned glial/biomatrix complexes direct neurite outgrowth. These data have significant implications for in vivo experiments focusing on glial transplantation. Transplanting glial/biomatrix complexes may stimulate the directional regrowth of severed axons across a lesion site.
Deumens
R
R
Department of Psychiatry and Neuropsychology, European Graduate School for Neuroscience (EURON), Universiteit Maastricht, PO Box 616, 6200 MD, Maastricht, The Netherlands. r.deumens@np.unimaas.nl
Koopmans
G C
GC
Den Bakker
C G J
CG
Maquet
V
V
Blacher
S
S
Honig
W M M
WM
Jérôme
R
R
Pirard
J-P
JP
Steinbusch
H W M
HW
Joosten
E A J
EA
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology; D016380:Brain Tissue Transplantation / Q000379:methods; D002450:Cell Communication / Q000502:physiology*; D016764:Cell Polarity / Q000502:physiology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000196:embryology / Q000502:physiology; D018920:Coculture Techniques / Q000379:methods; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D020439:Growth Cones / Q000502:physiology* / Q000648:ultrastructure; D007797:Laminin; D008297:Male; D009416:Nerve Regeneration / Q000502:physiology*; D016501:Neurites / Q000502:physiology / Q000648:ultrastructure; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009830:Olfactory Bulb / Q000166:cytology / Q000196:embryology / Q000502:physiology; D051381:Rats; D011917:Rats, Inbred Lew
C107909:Biomatrix preparation; D007797:Laminin