摘要
Objective To hibernate fetal ventral mesencephalic tegmental (VMT) cells from Spraque Dawley rats E 15 for 5 days at 4℃ 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 μg/ml), recombinant human glial cell derived neurotrophic factor (rhGDNF 20 μg/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 (F 17,1 =4.677; P= 0.045). After 5 days of hibernation, the viability of fetal VMT cells reduced by 30% (F 7,1 =88.493; P<0.001) in HM. However, THir cell density reduced by more than 90% as compared to the freshly harvested VMT cells (F 7,1 =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). Conclusion Supplementation of appropriate trophic factors for hibernated fetal VMT cells promotes cell viability and the subsequent expression of THir cell density.
Objective To hibernate fetal ventral mesencephalic tegmental (VMT) cells from Spraque Dawley rats E 15 for 5 days at 4℃ 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 μg/ml), recombinant human glial cell derived neurotrophic factor (rhGDNF 20 μg/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 (F 17,1 =4.677; P= 0.045). After 5 days of hibernation, the viability of fetal VMT cells reduced by 30% (F 7,1 =88.493; P<0.001) in HM. However, THir cell density reduced by more than 90% as compared to the freshly harvested VMT cells (F 7,1 =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). Conclusion Supplementation of appropriate trophic factors for hibernated fetal VMT cells promotes cell viability and the subsequent expression of THir cell density.
