TY - JOUR
T1 - Rat fetal ventral mesencephalon grown as solid tissue cultures
T2 - Influence of culture time and BDNF treatment on dopamine neuron survival and function
AU - Höglinger, Günter U.
AU - Sautter, Jürgen
AU - Meyer, Morten
AU - Spenger, Christian
AU - Seiler, Rolf W.
AU - Oertel, Wolfgang H.
AU - Widmer, Hans R.
N1 - Funding Information:
We thank Beatrice Bühler, Tanja Bosnjak, and Matthias Bischof for excellent technical assistance. This research was supported by the Swiss National Science Foundation (Grants No. 31-45558.95 and No. 31-52947.97), the Swiss Federal Office for Education and Science (Grant No. BBW 93.0349), and Desirée and Niels Ydes Foundation, Denmark. BDNF was kindly provided by Regeneron Pharmaceuticals, NY, USA.
PY - 1998/12/7
Y1 - 1998/12/7
N2 - Free-floating roller tube (FFRT) cultures of fetal rat and human nigral tissue are a means for tissue storage prior to grafting in experimental Parkinson's disease. In the present study, FFRT cultures prepared from embryonic-day-14 rat ventral mesencephalon were maintained for 4, 8, 12, or 16 days in vitro (DIV) in the presence or absence (controls) of BDNF [100 ng/ml]. The dopamine content in the culture medium, analyzed by HPLC, was significantly higher (4-5 fold) in the BDNF group at DIV 8 and DIV 12 compared to the corresponding control levels (40 pg/ml). The number of tyrosine hydroxylase immunoreactive neurons was significantly higher for BDNF treated cultures (2729 + 300) at DIV 8, as compared to controls (1679 ± 217). At DIV 12, the culture volume was significantly increased by BDNF (1.05 ±0.12 vs. 0.71 ± 0.04 mm3). Similar results were obtained for total protein. Western blot analysis demonstrated increasing signals for GFAP with increasing time in culture, but levels for control and BDNF treated cultures did not differ at any time-point investigated. In conclusion, it is suggested that the time window for effective storage of dopaminergic tissue prior to grafting can be extended by using the FFRT culture technique and that the in vitro storage may be further prolonged by treatment with BDNF.
AB - Free-floating roller tube (FFRT) cultures of fetal rat and human nigral tissue are a means for tissue storage prior to grafting in experimental Parkinson's disease. In the present study, FFRT cultures prepared from embryonic-day-14 rat ventral mesencephalon were maintained for 4, 8, 12, or 16 days in vitro (DIV) in the presence or absence (controls) of BDNF [100 ng/ml]. The dopamine content in the culture medium, analyzed by HPLC, was significantly higher (4-5 fold) in the BDNF group at DIV 8 and DIV 12 compared to the corresponding control levels (40 pg/ml). The number of tyrosine hydroxylase immunoreactive neurons was significantly higher for BDNF treated cultures (2729 + 300) at DIV 8, as compared to controls (1679 ± 217). At DIV 12, the culture volume was significantly increased by BDNF (1.05 ±0.12 vs. 0.71 ± 0.04 mm3). Similar results were obtained for total protein. Western blot analysis demonstrated increasing signals for GFAP with increasing time in culture, but levels for control and BDNF treated cultures did not differ at any time-point investigated. In conclusion, it is suggested that the time window for effective storage of dopaminergic tissue prior to grafting can be extended by using the FFRT culture technique and that the in vitro storage may be further prolonged by treatment with BDNF.
KW - Brain-derived neurotrophic factor
KW - Dopamine
KW - Mesencephalic tissue culture
KW - Parkinson's disease
UR - http://www.scopus.com/inward/record.url?scp=17144448554&partnerID=8YFLogxK
U2 - 10.1016/S0006-8993(98)01030-0
DO - 10.1016/S0006-8993(98)01030-0
M3 - Article
C2 - 9838177
AN - SCOPUS:17144448554
SN - 0006-8993
VL - 813
SP - 313
EP - 322
JO - Brain Research
JF - Brain Research
IS - 2
ER -
