Neurotrophic factors increase tyrosine hydroxylase immunoreactive cell density of fetal mesencephalic cells.
OBJECTIVE
To hibernate fetal ventral mesencephalic tegmental (VMT) cells from Spraque-Dawley rats E-15 for 5 days at 4 degrees C in either hibernation media (HM) or in conditioned hibernation media (CHM) supplemented with trophic factors such as epidermal growth factor (EGF 200 ng/ml), basic fibroblast growth factor (bFGF 100 ng/ml), recombinant human brain derived neurotrophic factor (rhBDNF 20 micrograms/ml), recombinant human glial cell-derived neurotrophic factor (rhGDNF 20 micrograms/ml), fetal calf serum (FCS 9%), or human placental cord serum (HPCS 10%).


METHODS
The percent of cell viability and the density of tyrosine hydroxylase immunoreactive (THir) cells in fetal striatal-VMT co-culture were determined.


RESULTS
The viability of fetal striatal cells (0.8 +/- 0.104) was slightly higher than that of fetal VMT cells (0.7 +/- 0.072) at 0-time point (F17,1 = 4.677; P = 0.045). After 5 days of hibernation, the viability of fetal VMT cells reduced by 30% (F7,1 = 88.493; P < 0.001) in HM. However, THir-cell density reduced by more than 90% as compared to the freshly harvested VMT cells (F7,1 = 179.944; P < 0.001). CHM with HPCS, bFGF, EGF, BDNF, and GDNF showed higher THir-cell density than that of HM or CHM supplemented with FCS (P < 0.001).


CONCLUSIONS
Supplementation of appropriate trophic factors for hibernated fetal VMT cells promotes cell viability and the subsequent expression of THir-cell density.
Thajeb
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Section of Neurology, Cathay General Hospital, Taipei, Tjioe Tjuan Neuroscience Research Laboratory, China.
D000818:Animals; D002452:Cell Count; D002469:Cell Separation; D002478:Cells, Cultured; D003080:Cold Temperature; D017077:Culture Media, Conditioned; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005333:Fetus; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013681:Tegmentum Mesencephali / Q000166:cytology* / Q000201:enzymology; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism*
D017077:Culture Media, Conditioned; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D014446:Tyrosine 3-Monooxygenase