Death in ventral mesenchephalic cell suspensions studied over time with electron microscope.

Death in ventral mesenchephalic cell suspensions studied over time with electron microscope.

Field: Disorders of the nervous system

Authors: Emgård, Mia
Brundin, Patrik

Address of presenting
author: Section for Neuronal Survival
Wallenberg Neuroscience Center
Sölvegatan 17
SE-223 62 Lund

E-mail: mia.emgard_mattson@mphy.lu.se

Phone: +46462220524

Fax: +46462220531

Text of abstract: Transplantation of dopaminergic neurons from embryonic ventral mesencephalon (VM) has been studied for more than 20 years with the aim of develping it as a treatment for Parkinson’ s disease. Several different approaches has been taken to increase the low survival of the grafted neurons, but even with the most effective treatment, only around 40% of the transplanted neurons survive. Most of the neurons die during the grafting procedure or within 4 days of implantation. In order to improve our understanding of cell death in transplants, the present study was undertaken to study the cell suspensions prior to transplantation.
Ventral mesencephali from rat embryos of embryonic day 14 were dissected out and prepared identically as for transplantation. The cell suspension was divided into 6 equal aliquots and were incubated for either 2 hours in room temperature, or 2 hours in room temperature followed by 2- or 6 hours in 37°C. Whole pieces of VM and cellsuspension taken directly after preparation were used as controls. After each incubation, cell viability was evaluated with trypan blue exclusion test. Cell suspensions were fixed with 0.5% glutaraldehyde in 1.5% paraformaldehyde, and thereafter prepared for electron microscopy.
Swollen mitochondria and outer membrane morphology was evaluated. Also, the amount of chromatin condensation and ruptured membranes and organells were evaluated. Signs of cellular damage was visable as swollen mitochindria and chromatin condensation, which indicates that there is an ongoing cell degeneration in the cell suspensions used for transplantation. We are finally aiming at investigating the effects of cyclosporin A and FK506 on the mitochondrial condition and cell viability.

Keywords: Parkinson's disease, transplantation, ventral mesencephalon, cell death, electron microscope

	Death in ventral mesenchephalic cell suspensions studied over time with electron microscope.
Field:	Disorders of the nervous system
Authors:	Emgård, Mia Brundin, Patrik
Address of presenting author:	Section for Neuronal Survival Wallenberg Neuroscience Center Sölvegatan 17 SE-223 62 Lund
E-mail:	mia.emgard_mattson@mphy.lu.se
Phone:	+46462220524
Fax:	+46462220531
Text of abstract:	Transplantation of dopaminergic neurons from embryonic ventral mesencephalon (VM) has been studied for more than 20 years with the aim of develping it as a treatment for Parkinson’ s disease. Several different approaches has been taken to increase the low survival of the grafted neurons, but even with the most effective treatment, only around 40% of the transplanted neurons survive. Most of the neurons die during the grafting procedure or within 4 days of implantation. In order to improve our understanding of cell death in transplants, the present study was undertaken to study the cell suspensions prior to transplantation. Ventral mesencephali from rat embryos of embryonic day 14 were dissected out and prepared identically as for transplantation. The cell suspension was divided into 6 equal aliquots and were incubated for either 2 hours in room temperature, or 2 hours in room temperature followed by 2- or 6 hours in 37°C. Whole pieces of VM and cellsuspension taken directly after preparation were used as controls. After each incubation, cell viability was evaluated with trypan blue exclusion test. Cell suspensions were fixed with 0.5% glutaraldehyde in 1.5% paraformaldehyde, and thereafter prepared for electron microscopy. Swollen mitochondria and outer membrane morphology was evaluated. Also, the amount of chromatin condensation and ruptured membranes and organells were evaluated. Signs of cellular damage was visable as swollen mitochindria and chromatin condensation, which indicates that there is an ongoing cell degeneration in the cell suspensions used for transplantation. We are finally aiming at investigating the effects of cyclosporin A and FK506 on the mitochondrial condition and cell viability.

Keywords:	Parkinson's disease, transplantation, ventral mesencephalon, cell death, electron microscope

Index

Created 2000-03-15

Department of Physiological Sciences, Lund University

Web programming FormOnLine AB