Abstract：
:Transplantation of human fetal ventral mesencephalon (VM) to Parkinsonian patients has shown beneficial effects in several clinical trials. However, further improvements in the transplantation technique are needed. Delayed surgery, i.e., the in vitro maintenance of the tissue prior to transplantation would present several advantages. The roller tube technique as initially described by Gähwiler (1981) was modified in several aspects for the long-term maintenance of dopaminergic neurones of human fetal VM. Tissue cultures were maintained free-floating in the medium for up to 42 days. The human fetal material was obtained from legal induced suction abortions. The embryonic age ranged from 5 to 12 weeks post-conception. Identification of VM was possible in 43% of the cases. Neurones in cultures were demonstrated by means of immunohistochemistry for tyrosine hydroxylase (TH) and gamma-amino butyric acid (GABA), by electron microscopy and by hybridisation histochemistry using a TH-mRNA-sensitive probe. A high variability in the number of TH-positive cells in individual cultures derived from the same embryo was observed. In 20 microns frozen sections of such tissue cultures the mean +/- SEM of TH-positive cells was 6.5 +/- 1.2/0.1 mm2 (n = 79; range: 0-73). The technique described insures the growth of long-term cultures of human fetal VM.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Spenger C,Studer L,Evtouchenko L,Egli M,Burgunder JM,Markwalder R,Seiler RWdoi
10.1016/0165-0270(94)90160-0subject
Has Abstractpub_date
1994-09-01 00:00:00pages
63-73issue
1eissn
0165-0270issn
1872-678Xpii
0165-0270(94)90160-0journal_volume
54pub_type
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