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Ultrastructural characterization of dissociated embryonic ventral mesencephalic tissue treated with neuroprotectants.

Ahn, Young Hwan LU ; Emgård-Mattson, Mia LU and Brundin, Patrik LU (2003) In Cell Transplantation 12(3). p.235-241
Abstract
Poor survival and differentiation of grafted dopamine neurons limits the application of clinical transplantation in Parkinson’s disease. The survival of grafted dopamine neurons is only improved by a factor of 2–3 by adding neuroprotectants during tissue preparation. We used dye exclusion cell viability and electron microscopy to investigate the effects of the caspase inhibitor ac-YVAD-cmk and the lazaroid tirilazad mesylate on ultrastructural changes in dissociated embryonic mesencephalic cells. In addition, we examined whether the neuroprotectants selectively counteracted specific signs of neurodegeneration. Cell viability decreased significantly over time in both control and treated cell suspensions, but the number of viable cells... (More)
Poor survival and differentiation of grafted dopamine neurons limits the application of clinical transplantation in Parkinson’s disease. The survival of grafted dopamine neurons is only improved by a factor of 2–3 by adding neuroprotectants during tissue preparation. We used dye exclusion cell viability and electron microscopy to investigate the effects of the caspase inhibitor ac-YVAD-cmk and the lazaroid tirilazad mesylate on ultrastructural changes in dissociated embryonic mesencephalic cells. In addition, we examined whether the neuroprotectants selectively counteracted specific signs of neurodegeneration. Cell viability decreased significantly over time in both control and treated cell suspensions, but the number of viable cells remaining was significantly higher in tirilazad mesylate-treated cell suspensions. In control samples, the proportion of cells with an ultrastructure consistent with healthy cells decreased from 70%, immediately after dissociation, to 30% after 8 h of incubation. Similar changes were also observed in cell suspensions treated with neuroprotectants. Thus, the neuroprotectants examined did not block the development of specific morphological signs of neurodegeneration. However, when also taking into account that dead cells lysed and disappeared from each cell suspension with time, we found that the total number of remaining viable cells with healthy nuclear chromatin or intact membrane integrity was significantly higher in the tirilazad mesylate-treated group. The results indicate that tirilazad mesylate protects only a small subpopulation of embryonic mesencephalic cells from degeneration induced by mechanical trauma during tissue dissection and dissociation. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Embryonic mesencephalic, Ultrastructure, Lazaroid, Caspase inhibitor, Viability, Survival
in
Cell Transplantation
volume
12
issue
3
pages
235 - 241
publisher
Cognizant Communication Corporation
external identifiers
  • pmid:12797378
  • wos:000182958900005
  • scopus:0038025359
ISSN
1555-3892
language
English
LU publication?
yes
id
420a2db4-e09f-449f-ae1f-292399bf809f (old id 116044)
alternative location
http://www.ingentaconnect.com/content/cog/ct/2003/00000012/00000003/ct388
date added to LUP
2007-06-26 08:50:16
date last changed
2018-01-07 06:15:06
@article{420a2db4-e09f-449f-ae1f-292399bf809f,
  abstract     = {Poor survival and differentiation of grafted dopamine neurons limits the application of clinical transplantation in Parkinson’s disease. The survival of grafted dopamine neurons is only improved by a factor of 2–3 by adding neuroprotectants during tissue preparation. We used dye exclusion cell viability and electron microscopy to investigate the effects of the caspase inhibitor ac-YVAD-cmk and the lazaroid tirilazad mesylate on ultrastructural changes in dissociated embryonic mesencephalic cells. In addition, we examined whether the neuroprotectants selectively counteracted specific signs of neurodegeneration. Cell viability decreased significantly over time in both control and treated cell suspensions, but the number of viable cells remaining was significantly higher in tirilazad mesylate-treated cell suspensions. In control samples, the proportion of cells with an ultrastructure consistent with healthy cells decreased from 70%, immediately after dissociation, to 30% after 8 h of incubation. Similar changes were also observed in cell suspensions treated with neuroprotectants. Thus, the neuroprotectants examined did not block the development of specific morphological signs of neurodegeneration. However, when also taking into account that dead cells lysed and disappeared from each cell suspension with time, we found that the total number of remaining viable cells with healthy nuclear chromatin or intact membrane integrity was significantly higher in the tirilazad mesylate-treated group. The results indicate that tirilazad mesylate protects only a small subpopulation of embryonic mesencephalic cells from degeneration induced by mechanical trauma during tissue dissection and dissociation.},
  author       = {Ahn, Young Hwan and Emgård-Mattson, Mia and Brundin, Patrik},
  issn         = {1555-3892},
  keyword      = {Embryonic mesencephalic,Ultrastructure,Lazaroid,Caspase inhibitor,Viability,Survival},
  language     = {eng},
  number       = {3},
  pages        = {235--241},
  publisher    = {Cognizant Communication Corporation},
  series       = {Cell Transplantation},
  title        = {Ultrastructural characterization of dissociated embryonic ventral mesencephalic tissue treated with neuroprotectants.},
  volume       = {12},
  year         = {2003},
}