| Publication number |
CLPUB00032 |
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| Authors |
Durand M.M., Chugani D.C., Mahmoudi M., Phelps M.E. |
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| Title |
Characterization of neuron-like cell line immortalized from primary rat mesencephalon cultures. |
|---|
| Citation |
Soc. Neurosci. Abstr. 16:40-40(1990) |
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| Web pages |
https://www.sfn.org/meetings/past-and-future-annual-meetings/1971-1999-abstracts |
|---|
| Abstract |
The transplantation of fetal mesencephalic tissue in striatum shows
promise as an effective treatment for Parkinson's disease. The limited
availability of human fetal tissue as well as complex ethical issues
regarding the use of the tissue will prevent widespread implementation of
this procedure. Reversibly immortalized mesencephalic dopaminergic cell
lines may be the answer to this problem. We have produced reversibly
immortalized cells from embryonic day 14 rat mesencephalon using a
retroviral vector containing the genes for temperature sensitive large T
antigen of SV40 and neomycin resistance (G418). This retrovirus was stably
produced by a psi 2 cell line obtained from R. McKay, MIT. Primary
cultures were enriched for dopaminergic cells by panning in tissue culture
plates that had been coated with the monoclonal antibody, neuron specific
protein 4. Following panning, cells were grown with supernatant from psi 2
cell line containing the retrovirus (dilution 1:3). After 48 hr the cells
were incubated with G418 to select for cells which had incorporated the
virus. Surviving cells, selected by dilution, proliferated at 33 Celsius
and displayed flat morphology. At 39 Celsius the cells stopped dividing and
some cells changed from flat cells with broad processes to neuron-like
morphology. One cell line CSM 14.1, expressed tyrosine hydroxylase and
neuron specific enolase immunoreactivity when raised to 39 Celsius for one
week. These cells were grown in the presence of various growth factors in
attempt to increase neuronal characteristics. The addition of basic
fibroblast growth factor produced greater than 90% neuron morphology after
one week in culture at 39 Celsius. These cells in addition expressed
neurofilament 68 and OL28 (a putative neuronal marker) immunoreactivity.
These data suggest that it may to possible to reversibly immortalize
mesencephalic dopaminergic cells for transplantation.
|
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| Cell lines |
CVCL_8182; CSM14.1 |
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