The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease
Exogenous cell replacement represents a potent treatment option for Parkinson's disease. However, the low survival rate of transplanted dopaminergic neurons (DA) calls for methodological improvements. Here we evaluated a method to combine transient genetic modification of neuronal progenitor ce...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
SAGE Publishing
2012-04-01
|
| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/096368911X586774 |
| _version_ | 1819019619524935680 |
|---|---|
| author | Andreas Ratzka Ieva Kalve Meltem Özer André Nobre Maike Wesemann Julia Jungnickel Christiane Köster-Patzlaff Olga Baron Claudia Grothe |
| author_facet | Andreas Ratzka Ieva Kalve Meltem Özer André Nobre Maike Wesemann Julia Jungnickel Christiane Köster-Patzlaff Olga Baron Claudia Grothe |
| author_sort | Andreas Ratzka |
| collection | DOAJ |
| description | Exogenous cell replacement represents a potent treatment option for Parkinson's disease. However, the low survival rate of transplanted dopaminergic neurons (DA) calls for methodological improvements. Here we evaluated a method to combine transient genetic modification of neuronal progenitor cells with an optimized cell culture protocol prior to intrastriatal transplantation into 6-hydroxydopamine (6-OHDA) unilateral lesioned rats. Plasmid-based delivery of brain-derived neurotrophic factor (BDNF) increases the number of DA neurons, identified by tyrosine hydroxylase immunoreactivity (TH-ir), by 25% in vitro, compared to enhanced green fluorescence protein (EGFP)-transfected controls. However, the nucleofection itself, especially the cell detachment and reseeding procedure, decreases the TH-ir neuron number to 40% compared with nontransfected control cultures. To circumvent this drawback we established the colayer method, which contains a mix of nucleofected cells reseeded on top of an adherent sister culture in a ratio 1:3. In this setup TH-ir neuron number remains high and could be further increased by 25% after BDNF transfection. Comparison of both cell culture procedures (standard and colayer) after intrastriatal transplantation revealed a similar DA neuron survival as seen in vitro. Two weeks after grafting TH-ir neuron number was strongly reduced in animals receiving the standard EGFP-transfected cells (271 ± 62) compared to 1,723 ± 199 TH-ir neurons in the colayer group. In contrast to the in vitro results, no differences in the number of grafted TH-ir neurons were observed between BDNF, EGFP, and nontransfected colayer groups, neither 2 nor 13 weeks after transplantation. Likewise, amphetamine and apomorphine-induced rotational behavior improved similarly over time in all groups. Nevertheless, the colayer protocol provides an efficient way for neurotrophic factor release by transplanted progenitor cells and will help to study the effects of candidate factors on survival and integration of transplanted DA neurons. |
| first_indexed | 2024-12-21T03:38:12Z |
| format | Article |
| id | doaj.art-acd627d973d84b49826a3bf3f95ef662 |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-21T03:38:12Z |
| publishDate | 2012-04-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-acd627d973d84b49826a3bf3f95ef6622022-12-21T19:17:17ZengSAGE PublishingCell Transplantation0963-68971555-38922012-04-012110.3727/096368911X586774The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's DiseaseAndreas Ratzka0Ieva Kalve1Meltem Özer2André Nobre3Maike Wesemann4Julia Jungnickel5Christiane Köster-Patzlaff6Olga Baron7Claudia Grothe8Institute of Neuroanatomy, Hannover Medical School, Hannover, GermanyCenter for Systems Neuroscience (ZSN), Hannover, GermanyCenter for Systems Neuroscience (ZSN), Hannover, GermanyCenter for Systems Neuroscience (ZSN), Hannover, GermanyInstitute of Neuroanatomy, Hannover Medical School, Hannover, GermanyInstitute of Neuroanatomy, Hannover Medical School, Hannover, GermanyInstitute of Neuroanatomy, Hannover Medical School, Hannover, GermanyCenter for Systems Neuroscience (ZSN), Hannover, GermanyCenter for Systems Neuroscience (ZSN), Hannover, GermanyExogenous cell replacement represents a potent treatment option for Parkinson's disease. However, the low survival rate of transplanted dopaminergic neurons (DA) calls for methodological improvements. Here we evaluated a method to combine transient genetic modification of neuronal progenitor cells with an optimized cell culture protocol prior to intrastriatal transplantation into 6-hydroxydopamine (6-OHDA) unilateral lesioned rats. Plasmid-based delivery of brain-derived neurotrophic factor (BDNF) increases the number of DA neurons, identified by tyrosine hydroxylase immunoreactivity (TH-ir), by 25% in vitro, compared to enhanced green fluorescence protein (EGFP)-transfected controls. However, the nucleofection itself, especially the cell detachment and reseeding procedure, decreases the TH-ir neuron number to 40% compared with nontransfected control cultures. To circumvent this drawback we established the colayer method, which contains a mix of nucleofected cells reseeded on top of an adherent sister culture in a ratio 1:3. In this setup TH-ir neuron number remains high and could be further increased by 25% after BDNF transfection. Comparison of both cell culture procedures (standard and colayer) after intrastriatal transplantation revealed a similar DA neuron survival as seen in vitro. Two weeks after grafting TH-ir neuron number was strongly reduced in animals receiving the standard EGFP-transfected cells (271 ± 62) compared to 1,723 ± 199 TH-ir neurons in the colayer group. In contrast to the in vitro results, no differences in the number of grafted TH-ir neurons were observed between BDNF, EGFP, and nontransfected colayer groups, neither 2 nor 13 weeks after transplantation. Likewise, amphetamine and apomorphine-induced rotational behavior improved similarly over time in all groups. Nevertheless, the colayer protocol provides an efficient way for neurotrophic factor release by transplanted progenitor cells and will help to study the effects of candidate factors on survival and integration of transplanted DA neurons.https://doi.org/10.3727/096368911X586774 |
| spellingShingle | Andreas Ratzka Ieva Kalve Meltem Özer André Nobre Maike Wesemann Julia Jungnickel Christiane Köster-Patzlaff Olga Baron Claudia Grothe The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease Cell Transplantation |
| title | The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease |
| title_full | The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease |
| title_fullStr | The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease |
| title_full_unstemmed | The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease |
| title_short | The Colayer Method as an Efficient Way to Genetically Modify Mesencephalic Progenitor Cells Transplanted into 6-OHDA Rat Model of Parkinson's Disease |
| title_sort | colayer method as an efficient way to genetically modify mesencephalic progenitor cells transplanted into 6 ohda rat model of parkinson s disease |
| url | https://doi.org/10.3727/096368911X586774 |
| work_keys_str_mv | AT andreasratzka thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT ievakalve thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT meltemozer thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT andrenobre thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT maikewesemann thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT juliajungnickel thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT christianekosterpatzlaff thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT olgabaron thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT claudiagrothe thecolayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT andreasratzka colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT ievakalve colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT meltemozer colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT andrenobre colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT maikewesemann colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT juliajungnickel colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT christianekosterpatzlaff colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT olgabaron colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease AT claudiagrothe colayermethodasanefficientwaytogeneticallymodifymesencephalicprogenitorcellstransplantedinto6ohdaratmodelofparkinsonsdisease |