Nucleofection Is the Most Efficient Nonviral Transfection Method for Neuronal Stem Cells Derived from Ventral Mesencephali with No Changes in Cell Composition or Dopaminergic Fate
Neuronal progenitor cells (NPCs) play an important role in potential regenerative therapeutic strategies for neurodegenerative diseases, such as Parkinson disease. However, survival of transplanted cells is, as yet, limited, and the identification of grafted cells in situ remains difficult. The use...
Gespeichert in:
| Veröffentlicht in: | Stem cells (Dayton, Ohio) Ohio), 2006-12, Vol.24 (12), p.2776-2791 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2791 |
|---|---|
| container_issue | 12 |
| container_start_page | 2776 |
| container_title | Stem cells (Dayton, Ohio) |
| container_volume | 24 |
| creator | Cesnulevicius, Konstantin Timmer, Marco Wesemann, Maike Thomas, Tobias Barkhausen, Tanja Grothe, Claudia |
| description | Neuronal progenitor cells (NPCs) play an important role in potential regenerative therapeutic strategies for neurodegenerative diseases, such as Parkinson disease. However, survival of transplanted cells is, as yet, limited, and the identification of grafted cells in situ remains difficult. The use of NPCs could be more effective with regard to a better survival and maturation when transfected with one or more neurotrophic factors. Therefore, we investigated the possibility of transfecting mesencephalic neuronal progenitors with different constructs carrying neurotrophic factors or the expression reporters enhanced green fluorescence protein (EGFP) and red fluorescent protein (DsRed). Different techniques for transfection were compared, and the highest transfection rate of up to 47% was achieved by nucleofection. Mesencephalic neuronal progenitors survived the transfection procedure; 6 hours after transfection, viability was approximately 40%, and the transfected cells differentiated into, for example, tyrosine hydroxylase‐positive neurons. Within the group of transfected cells, many progenitors and several neurons were found. To provide the progenitor cells with a neurotrophic factor, different isoforms of fibroblast growth factor‐2 were introduced. To follow the behavior of the transfected cells in vitro, functional tests such as the cell viability assay (water‐soluble tetrazolium salt assay [WST‐1]) and the cell proliferation assay (5‐bromo‐2′‐deoxyuridine‐enzyme‐linked immunosorbent assay) were performed. In addition, these transfected NPCs were viable after transplantation, expressed tyrosine hydroxylase in vivo, and could easily be detected within the host striatum because of their EGFP expression. This study shows that genetic modification of neural progenitors could provide attractive perspectives for new therapeutic concepts in neurodegenerative diseases. |
| doi_str_mv | 10.1634/stemcells.2006-0176 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_68217334</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>68217334</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4316-97b689fa59d4e7f248007febbfd6831631490491553b8ff0fa485692cf0f39363</originalsourceid><addsrcrecordid>eNqNkU1v1DAQhiMEol_8AiTkE7cUO_5ILE4o3dJK3e2hC9fIyY4bo8QOttOqv6t_EKe7wBFOHsnPPDOaN8veE3xOBGWfQoSxg2EI5wXGIsekFK-yY8KZzJkk1etUYyFyjqU8yk5C-IExYbyq3mZHREhcECmOs-fN3A3gNHTROIuuA4o9oLULEa20Np0BG9HG2Qfj1YC2Xtnwm11D7N0OaefRBmbvbALu0k6oXpZCF-DNA6R_70b0PWkWwRoC2A6mXg0GPZrYJzeqe2XvISBjX1pR7cbJBfMyJMkv3KRGY8Hfmw5dqghn2RuthgDvDu9p9u1yta2v8pvbr9f1l5u8Y5SIXJatqKRWXO4YlLpgFcalhrbVO1ElgBImcboU57SttMZasYoLWXSppJIKepp93Hsn737OEGIzmrBcXFlwc2hEVZCSUvZPkEhepEk8gXQPdt6F4EE3kzej8k8Nwc0SavMn1GYJtVlCTV0fDvq5HWH3t-eQYgI-74FHM8DT_zibu-1qXbCiTPpf0nG1ZA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>19520495</pqid></control><display><type>article</type><title>Nucleofection Is the Most Efficient Nonviral Transfection Method for Neuronal Stem Cells Derived from Ventral Mesencephali with No Changes in Cell Composition or Dopaminergic Fate</title><source>MEDLINE</source><source>Oxford University Press Journals All Titles (1996-Current)</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Cesnulevicius, Konstantin ; Timmer, Marco ; Wesemann, Maike ; Thomas, Tobias ; Barkhausen, Tanja ; Grothe, Claudia</creator><creatorcontrib>Cesnulevicius, Konstantin ; Timmer, Marco ; Wesemann, Maike ; Thomas, Tobias ; Barkhausen, Tanja ; Grothe, Claudia</creatorcontrib><description>Neuronal progenitor cells (NPCs) play an important role in potential regenerative therapeutic strategies for neurodegenerative diseases, such as Parkinson disease. However, survival of transplanted cells is, as yet, limited, and the identification of grafted cells in situ remains difficult. The use of NPCs could be more effective with regard to a better survival and maturation when transfected with one or more neurotrophic factors. Therefore, we investigated the possibility of transfecting mesencephalic neuronal progenitors with different constructs carrying neurotrophic factors or the expression reporters enhanced green fluorescence protein (EGFP) and red fluorescent protein (DsRed). Different techniques for transfection were compared, and the highest transfection rate of up to 47% was achieved by nucleofection. Mesencephalic neuronal progenitors survived the transfection procedure; 6 hours after transfection, viability was approximately 40%, and the transfected cells differentiated into, for example, tyrosine hydroxylase‐positive neurons. Within the group of transfected cells, many progenitors and several neurons were found. To provide the progenitor cells with a neurotrophic factor, different isoforms of fibroblast growth factor‐2 were introduced. To follow the behavior of the transfected cells in vitro, functional tests such as the cell viability assay (water‐soluble tetrazolium salt assay [WST‐1]) and the cell proliferation assay (5‐bromo‐2′‐deoxyuridine‐enzyme‐linked immunosorbent assay) were performed. In addition, these transfected NPCs were viable after transplantation, expressed tyrosine hydroxylase in vivo, and could easily be detected within the host striatum because of their EGFP expression. This study shows that genetic modification of neural progenitors could provide attractive perspectives for new therapeutic concepts in neurodegenerative diseases.</description><identifier>ISSN: 1066-5099</identifier><identifier>EISSN: 1549-4918</identifier><identifier>DOI: 10.1634/stemcells.2006-0176</identifier><identifier>PMID: 16902196</identifier><language>eng</language><publisher>Bristol: John Wiley & Sons, Ltd</publisher><subject>Animals ; Cell Count ; Cell Differentiation - drug effects ; Cell Lineage ; Cell Nucleus - drug effects ; Cell Nucleus - metabolism ; Cell Survival - drug effects ; Dopamine - metabolism ; Dopaminergic neurons ; Electroporation ; Fibroblast Growth Factor 2 - pharmacology ; Flow Cytometry ; Green Fluorescent Proteins - metabolism ; Mesencephalon - cytology ; Mesencephalon - pathology ; Mesencephalon - transplantation ; Neural stem cells ; Neurons - cytology ; Nucleofection ; Oxidopamine - pharmacology ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; Stem Cells - cytology ; Transfection ; Transfection - methods ; Transplantation ; Viruses</subject><ispartof>Stem cells (Dayton, Ohio), 2006-12, Vol.24 (12), p.2776-2791</ispartof><rights>Copyright © 2006 AlphaMed Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4316-97b689fa59d4e7f248007febbfd6831631490491553b8ff0fa485692cf0f39363</citedby><cites>FETCH-LOGICAL-c4316-97b689fa59d4e7f248007febbfd6831631490491553b8ff0fa485692cf0f39363</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,27906,27907</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16902196$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cesnulevicius, Konstantin</creatorcontrib><creatorcontrib>Timmer, Marco</creatorcontrib><creatorcontrib>Wesemann, Maike</creatorcontrib><creatorcontrib>Thomas, Tobias</creatorcontrib><creatorcontrib>Barkhausen, Tanja</creatorcontrib><creatorcontrib>Grothe, Claudia</creatorcontrib><title>Nucleofection Is the Most Efficient Nonviral Transfection Method for Neuronal Stem Cells Derived from Ventral Mesencephali with No Changes in Cell Composition or Dopaminergic Fate</title><title>Stem cells (Dayton, Ohio)</title><addtitle>Stem Cells</addtitle><description>Neuronal progenitor cells (NPCs) play an important role in potential regenerative therapeutic strategies for neurodegenerative diseases, such as Parkinson disease. However, survival of transplanted cells is, as yet, limited, and the identification of grafted cells in situ remains difficult. The use of NPCs could be more effective with regard to a better survival and maturation when transfected with one or more neurotrophic factors. Therefore, we investigated the possibility of transfecting mesencephalic neuronal progenitors with different constructs carrying neurotrophic factors or the expression reporters enhanced green fluorescence protein (EGFP) and red fluorescent protein (DsRed). Different techniques for transfection were compared, and the highest transfection rate of up to 47% was achieved by nucleofection. Mesencephalic neuronal progenitors survived the transfection procedure; 6 hours after transfection, viability was approximately 40%, and the transfected cells differentiated into, for example, tyrosine hydroxylase‐positive neurons. Within the group of transfected cells, many progenitors and several neurons were found. To provide the progenitor cells with a neurotrophic factor, different isoforms of fibroblast growth factor‐2 were introduced. To follow the behavior of the transfected cells in vitro, functional tests such as the cell viability assay (water‐soluble tetrazolium salt assay [WST‐1]) and the cell proliferation assay (5‐bromo‐2′‐deoxyuridine‐enzyme‐linked immunosorbent assay) were performed. In addition, these transfected NPCs were viable after transplantation, expressed tyrosine hydroxylase in vivo, and could easily be detected within the host striatum because of their EGFP expression. This study shows that genetic modification of neural progenitors could provide attractive perspectives for new therapeutic concepts in neurodegenerative diseases.</description><subject>Animals</subject><subject>Cell Count</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Lineage</subject><subject>Cell Nucleus - drug effects</subject><subject>Cell Nucleus - metabolism</subject><subject>Cell Survival - drug effects</subject><subject>Dopamine - metabolism</subject><subject>Dopaminergic neurons</subject><subject>Electroporation</subject><subject>Fibroblast Growth Factor 2 - pharmacology</subject><subject>Flow Cytometry</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Mesencephalon - cytology</subject><subject>Mesencephalon - pathology</subject><subject>Mesencephalon - transplantation</subject><subject>Neural stem cells</subject><subject>Neurons - cytology</subject><subject>Nucleofection</subject><subject>Oxidopamine - pharmacology</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Stem Cell Transplantation</subject><subject>Stem Cells - cytology</subject><subject>Transfection</subject><subject>Transfection - methods</subject><subject>Transplantation</subject><subject>Viruses</subject><issn>1066-5099</issn><issn>1549-4918</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1v1DAQhiMEol_8AiTkE7cUO_5ILE4o3dJK3e2hC9fIyY4bo8QOttOqv6t_EKe7wBFOHsnPPDOaN8veE3xOBGWfQoSxg2EI5wXGIsekFK-yY8KZzJkk1etUYyFyjqU8yk5C-IExYbyq3mZHREhcECmOs-fN3A3gNHTROIuuA4o9oLULEa20Np0BG9HG2Qfj1YC2Xtnwm11D7N0OaefRBmbvbALu0k6oXpZCF-DNA6R_70b0PWkWwRoC2A6mXg0GPZrYJzeqe2XvISBjX1pR7cbJBfMyJMkv3KRGY8Hfmw5dqghn2RuthgDvDu9p9u1yta2v8pvbr9f1l5u8Y5SIXJatqKRWXO4YlLpgFcalhrbVO1ElgBImcboU57SttMZasYoLWXSppJIKepp93Hsn737OEGIzmrBcXFlwc2hEVZCSUvZPkEhepEk8gXQPdt6F4EE3kzej8k8Nwc0SavMn1GYJtVlCTV0fDvq5HWH3t-eQYgI-74FHM8DT_zibu-1qXbCiTPpf0nG1ZA</recordid><startdate>200612</startdate><enddate>200612</enddate><creator>Cesnulevicius, Konstantin</creator><creator>Timmer, Marco</creator><creator>Wesemann, Maike</creator><creator>Thomas, Tobias</creator><creator>Barkhausen, Tanja</creator><creator>Grothe, Claudia</creator><general>John Wiley & Sons, Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>200612</creationdate><title>Nucleofection Is the Most Efficient Nonviral Transfection Method for Neuronal Stem Cells Derived from Ventral Mesencephali with No Changes in Cell Composition or Dopaminergic Fate</title><author>Cesnulevicius, Konstantin ; Timmer, Marco ; Wesemann, Maike ; Thomas, Tobias ; Barkhausen, Tanja ; Grothe, Claudia</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4316-97b689fa59d4e7f248007febbfd6831631490491553b8ff0fa485692cf0f39363</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Animals</topic><topic>Cell Count</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Lineage</topic><topic>Cell Nucleus - drug effects</topic><topic>Cell Nucleus - metabolism</topic><topic>Cell Survival - drug effects</topic><topic>Dopamine - metabolism</topic><topic>Dopaminergic neurons</topic><topic>Electroporation</topic><topic>Fibroblast Growth Factor 2 - pharmacology</topic><topic>Flow Cytometry</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>Mesencephalon - cytology</topic><topic>Mesencephalon - pathology</topic><topic>Mesencephalon - transplantation</topic><topic>Neural stem cells</topic><topic>Neurons - cytology</topic><topic>Nucleofection</topic><topic>Oxidopamine - pharmacology</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Stem Cell Transplantation</topic><topic>Stem Cells - cytology</topic><topic>Transfection</topic><topic>Transfection - methods</topic><topic>Transplantation</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cesnulevicius, Konstantin</creatorcontrib><creatorcontrib>Timmer, Marco</creatorcontrib><creatorcontrib>Wesemann, Maike</creatorcontrib><creatorcontrib>Thomas, Tobias</creatorcontrib><creatorcontrib>Barkhausen, Tanja</creatorcontrib><creatorcontrib>Grothe, Claudia</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Stem cells (Dayton, Ohio)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cesnulevicius, Konstantin</au><au>Timmer, Marco</au><au>Wesemann, Maike</au><au>Thomas, Tobias</au><au>Barkhausen, Tanja</au><au>Grothe, Claudia</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Nucleofection Is the Most Efficient Nonviral Transfection Method for Neuronal Stem Cells Derived from Ventral Mesencephali with No Changes in Cell Composition or Dopaminergic Fate</atitle><jtitle>Stem cells (Dayton, Ohio)</jtitle><addtitle>Stem Cells</addtitle><date>2006-12</date><risdate>2006</risdate><volume>24</volume><issue>12</issue><spage>2776</spage><epage>2791</epage><pages>2776-2791</pages><issn>1066-5099</issn><eissn>1549-4918</eissn><abstract>Neuronal progenitor cells (NPCs) play an important role in potential regenerative therapeutic strategies for neurodegenerative diseases, such as Parkinson disease. However, survival of transplanted cells is, as yet, limited, and the identification of grafted cells in situ remains difficult. The use of NPCs could be more effective with regard to a better survival and maturation when transfected with one or more neurotrophic factors. Therefore, we investigated the possibility of transfecting mesencephalic neuronal progenitors with different constructs carrying neurotrophic factors or the expression reporters enhanced green fluorescence protein (EGFP) and red fluorescent protein (DsRed). Different techniques for transfection were compared, and the highest transfection rate of up to 47% was achieved by nucleofection. Mesencephalic neuronal progenitors survived the transfection procedure; 6 hours after transfection, viability was approximately 40%, and the transfected cells differentiated into, for example, tyrosine hydroxylase‐positive neurons. Within the group of transfected cells, many progenitors and several neurons were found. To provide the progenitor cells with a neurotrophic factor, different isoforms of fibroblast growth factor‐2 were introduced. To follow the behavior of the transfected cells in vitro, functional tests such as the cell viability assay (water‐soluble tetrazolium salt assay [WST‐1]) and the cell proliferation assay (5‐bromo‐2′‐deoxyuridine‐enzyme‐linked immunosorbent assay) were performed. In addition, these transfected NPCs were viable after transplantation, expressed tyrosine hydroxylase in vivo, and could easily be detected within the host striatum because of their EGFP expression. This study shows that genetic modification of neural progenitors could provide attractive perspectives for new therapeutic concepts in neurodegenerative diseases.</abstract><cop>Bristol</cop><pub>John Wiley & Sons, Ltd</pub><pmid>16902196</pmid><doi>10.1634/stemcells.2006-0176</doi><tpages>16</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1066-5099 |
| ispartof | Stem cells (Dayton, Ohio), 2006-12, Vol.24 (12), p.2776-2791 |
| issn | 1066-5099 1549-4918 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_68217334 |
| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Animals Cell Count Cell Differentiation - drug effects Cell Lineage Cell Nucleus - drug effects Cell Nucleus - metabolism Cell Survival - drug effects Dopamine - metabolism Dopaminergic neurons Electroporation Fibroblast Growth Factor 2 - pharmacology Flow Cytometry Green Fluorescent Proteins - metabolism Mesencephalon - cytology Mesencephalon - pathology Mesencephalon - transplantation Neural stem cells Neurons - cytology Nucleofection Oxidopamine - pharmacology Rats Rats, Sprague-Dawley Stem Cell Transplantation Stem Cells - cytology Transfection Transfection - methods Transplantation Viruses |
| title | Nucleofection Is the Most Efficient Nonviral Transfection Method for Neuronal Stem Cells Derived from Ventral Mesencephali with No Changes in Cell Composition or Dopaminergic Fate |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-17T10%3A23%3A42IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Nucleofection%20Is%20the%20Most%20Efficient%20Nonviral%20Transfection%20Method%20for%20Neuronal%20Stem%20Cells%20Derived%20from%20Ventral%20Mesencephali%20with%20No%20Changes%20in%20Cell%20Composition%20or%20Dopaminergic%20Fate&rft.jtitle=Stem%20cells%20(Dayton,%20Ohio)&rft.au=Cesnulevicius,%20Konstantin&rft.date=2006-12&rft.volume=24&rft.issue=12&rft.spage=2776&rft.epage=2791&rft.pages=2776-2791&rft.issn=1066-5099&rft.eissn=1549-4918&rft_id=info:doi/10.1634/stemcells.2006-0176&rft_dat=%3Cproquest_cross%3E68217334%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=19520495&rft_id=info:pmid/16902196&rfr_iscdi=true |