In vitro characterization of trophic factor expression in neural precursor cells.
In cellular transplantation strategies for repairing the injured central nervous system, interactions between transplanted neural precursor cells (NPCs) and host tissue remain incompletely understood. Although trophins may contribute to the benefits observed, little research has explored this possibility. Candidate trophic factors were identified, and primers were designed for these genes. Template RNA was isolated from 3 NPC sources, and also from bone marrow stromal cells (BMSCs) and embryonic fibroblasts as comparative controls. Quantitative polymerase chain reaction was performed to determine the effect of cell source, passaging, cellular differentiation, and environmental changes on trophin factor expression in NPCs. Results were analyzed with multivariate statistical analyses. NPCs, BMSCs, and fibroblasts each expressed trophic factors in unique patterns. Trophic factor expression was similar among NPCs whether harvested from rat or mouse, brain or spinal cord, or their time in culture. The expression of neurotrophin NT-3, NT-4/5, glial-derived neurotrophic factor, and insulin-like growth factor-1 decreased with time in culture. Induced differentiation of NPCs led to a marked and statistically significant increase in the expression of trophic factors. Culture conditions and environmental changes were also associated with significant changes in trophin expression. These results suggest that trophins could contribute to the benefits associated with transplantation of NPCs as well as BMSCs. Trophic factor expression changes with NPC differentiation and environmental conditions, which could have important implications with regard to their behavior after in vivo transplantation.
Division of Genetics and Development, Krembil Neuroscience Centre, Toronto Western Research Institute, University Health Network, Toronto, Ontario, Canada.
Gregory W J
GW
Hawryluk
Andrea J
AJ
Mothe
Mahmood
M
Chamankhah
Jian
J
Wang
Charles
C
Tator
Michael G
MG
Fehlings
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D003470:Culture Media / Q000378:metabolism; D061252:Feeder Cells; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D013116:Spinal Cord / Q000166:cytology* / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D009414:Nerve Growth Factors; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; C518911:neurotropin 3, mouse; C467480:vascular endothelial growth factor A, mouse; D049452:Green Fluorescent Proteins; D012313:RNA; D007334:Insulin-Like Growth Factor I; C073922:neurotrophin 4