Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for Dopamine Neurons From Human Pluripotent Stem Cells.
Identifieur interne : 000934 ( Main/Exploration ); précédent : 000933; suivant : 000935Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for Dopamine Neurons From Human Pluripotent Stem Cells.
Auteurs : Julio Cesar Aguila [États-Unis] ; Alexandra Blak [États-Unis] ; Joris Van Arensbergen [États-Unis] ; Amaia Sousa [États-Unis] ; Nerea Vázquez [États-Unis] ; Ariane Aduriz [États-Unis] ; Mayela Gayosso [États-Unis] ; Maria Paz Lopez Mato [États-Unis] ; Rakel Lopez De Maturana [États-Unis] ; Eva Hedlund [États-Unis] ; Kai-Christian Sonntag [États-Unis] ; Rosario Sanchez-PernauteSource :
- Stem cells translational medicine [ 2157-6564 ] ; 2014.
English descriptors
- KwdEn :
- Blotting, Western, Cell Separation (methods), Dopaminergic Neurons (cytology), Flow Cytometry, Fluorescent Antibody Technique, Genetic Vectors, Green Fluorescent Proteins (genetics), Hepatocyte Nuclear Factor 3-beta (genetics), Humans, Lentivirus, Microscopy, Confocal, Neural Stem Cells (cytology), Pluripotent Stem Cells (cytology), Promoter Regions, Genetic, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Transduction, Genetic.
- MESH :
- chemical , genetics : Green Fluorescent Proteins, Hepatocyte Nuclear Factor 3-beta.
- cytology : Dopaminergic Neurons, Neural Stem Cells, Pluripotent Stem Cells.
- methods : Cell Separation.
- Blotting, Western, Flow Cytometry, Fluorescent Antibody Technique, Genetic Vectors, Humans, Lentivirus, Microscopy, Confocal, Promoter Regions, Genetic, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Transduction, Genetic.
Abstract
Human embryonic and induced pluripotent stem cells are potential cell sources for regenerative approaches in Parkinson disease. Inductive differentiation protocols can generate midbrain dopamine neurons but result in heterogeneous cell mixtures. Therefore, selection strategies are necessary to obtain uniform dopamine cell populations. Here, we developed a selection approach using lentivirus vectors to express green fluorescent protein under the promoter region of FOXA2, a transcription factor that is expressed in the floor plate domain that gives rise to dopamine neurons during embryogenesis. We first validated the specificity of the vectors in human cell lines against a promoterless construct. We then selected FOXA2-positive neural progenitors from several human pluripotent stem cell lines, which demonstrated a gene expression profile typical for the ventral domain of the midbrain and floor plate, but failed to enrich for dopamine neurons. To investigate whether this was due to the selection approach, we overexpressed FOXA2 in neural progenitors derived from human pluripotent stem cell lines. FOXA2 forced expression resulted in an increased expression of floor plate but not mature neuronal markers. Furthermore, selection of the FOXA2 overexpressing fraction also failed to enrich for dopamine neurons. Collectively, our results suggest that FOXA2 is not sufficient to induce a dopaminergic fate in this system. On the other hand, our study demonstrates that a combined approach of promoter activation and lentivirus vector technology can be used as a versatile tool for the selection of a defined cell population from a variety of human pluripotent stem cell lines.
DOI: 10.5966/sctm.2014-0011
PubMed: 25024431
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 000692
- to stream PubMed, to step Curation: 000692
- to stream PubMed, to step Checkpoint: 000692
- to stream Ncbi, to step Merge: 001827
- to stream Ncbi, to step Curation: 001827
- to stream Ncbi, to step Checkpoint: 001827
- to stream Main, to step Merge: 000936
- to stream Main, to step Curation: 000934
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for Dopamine Neurons From Human Pluripotent Stem Cells.</title><author><name sortKey="Aguila, Julio Cesar" sort="Aguila, Julio Cesar" uniqKey="Aguila J" first="Julio Cesar" last="Aguila">Julio Cesar Aguila</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Blak, Alexandra" sort="Blak, Alexandra" uniqKey="Blak A" first="Alexandra" last="Blak">Alexandra Blak</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Van Arensbergen, Joris" sort="Van Arensbergen, Joris" uniqKey="Van Arensbergen J" first="Joris" last="Van Arensbergen">Joris Van Arensbergen</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sousa, Amaia" sort="Sousa, Amaia" uniqKey="Sousa A" first="Amaia" last="Sousa">Amaia Sousa</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Vazquez, Nerea" sort="Vazquez, Nerea" uniqKey="Vazquez N" first="Nerea" last="Vázquez">Nerea Vázquez</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Aduriz, Ariane" sort="Aduriz, Ariane" uniqKey="Aduriz A" first="Ariane" last="Aduriz">Ariane Aduriz</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Gayosso, Mayela" sort="Gayosso, Mayela" uniqKey="Gayosso M" first="Mayela" last="Gayosso">Mayela Gayosso</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Lopez Mato, Maria Paz" sort="Lopez Mato, Maria Paz" uniqKey="Lopez Mato M" first="Maria Paz" last="Lopez Mato">Maria Paz Lopez Mato</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Lopez De Maturana, Rakel" sort="Lopez De Maturana, Rakel" uniqKey="Lopez De Maturana R" first="Rakel" last="Lopez De Maturana">Rakel Lopez De Maturana</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Hedlund, Eva" sort="Hedlund, Eva" uniqKey="Hedlund E" first="Eva" last="Hedlund">Eva Hedlund</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sonntag, Kai Christian" sort="Sonntag, Kai Christian" uniqKey="Sonntag K" first="Kai-Christian" last="Sonntag">Kai-Christian Sonntag</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sanchez Pernaute, Rosario" sort="Sanchez Pernaute, Rosario" uniqKey="Sanchez Pernaute R" first="Rosario" last="Sanchez-Pernaute">Rosario Sanchez-Pernaute</name><affiliation><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA rpernaute@inbiomed.org.</nlm:affiliation><wicri:noCountry code="subField">USA rpernaute@inbiomed.org.</wicri:noCountry></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2014">2014</date><idno type="RBID">pubmed:25024431</idno><idno type="pmid">25024431</idno><idno type="doi">10.5966/sctm.2014-0011</idno><idno type="wicri:Area/PubMed/Corpus">000692</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000692</idno><idno type="wicri:Area/PubMed/Curation">000692</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000692</idno><idno type="wicri:Area/PubMed/Checkpoint">000692</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000692</idno><idno type="wicri:Area/Ncbi/Merge">001827</idno><idno type="wicri:Area/Ncbi/Curation">001827</idno><idno type="wicri:Area/Ncbi/Checkpoint">001827</idno><idno type="wicri:doubleKey">2157-6564:2014:Aguila J:selection:based:on</idno><idno type="wicri:Area/Main/Merge">000936</idno><idno type="wicri:Area/Main/Curation">000934</idno><idno type="wicri:Area/Main/Exploration">000934</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for Dopamine Neurons From Human Pluripotent Stem Cells.</title><author><name sortKey="Aguila, Julio Cesar" sort="Aguila, Julio Cesar" uniqKey="Aguila J" first="Julio Cesar" last="Aguila">Julio Cesar Aguila</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Blak, Alexandra" sort="Blak, Alexandra" uniqKey="Blak A" first="Alexandra" last="Blak">Alexandra Blak</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Van Arensbergen, Joris" sort="Van Arensbergen, Joris" uniqKey="Van Arensbergen J" first="Joris" last="Van Arensbergen">Joris Van Arensbergen</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sousa, Amaia" sort="Sousa, Amaia" uniqKey="Sousa A" first="Amaia" last="Sousa">Amaia Sousa</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Vazquez, Nerea" sort="Vazquez, Nerea" uniqKey="Vazquez N" first="Nerea" last="Vázquez">Nerea Vázquez</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Aduriz, Ariane" sort="Aduriz, Ariane" uniqKey="Aduriz A" first="Ariane" last="Aduriz">Ariane Aduriz</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Gayosso, Mayela" sort="Gayosso, Mayela" uniqKey="Gayosso M" first="Mayela" last="Gayosso">Mayela Gayosso</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Lopez Mato, Maria Paz" sort="Lopez Mato, Maria Paz" uniqKey="Lopez Mato M" first="Maria Paz" last="Lopez Mato">Maria Paz Lopez Mato</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Lopez De Maturana, Rakel" sort="Lopez De Maturana, Rakel" uniqKey="Lopez De Maturana R" first="Rakel" last="Lopez De Maturana">Rakel Lopez De Maturana</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Hedlund, Eva" sort="Hedlund, Eva" uniqKey="Hedlund E" first="Eva" last="Hedlund">Eva Hedlund</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sonntag, Kai Christian" sort="Sonntag, Kai Christian" uniqKey="Sonntag K" first="Kai-Christian" last="Sonntag">Kai-Christian Sonntag</name><affiliation wicri:level="2"><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sanchez Pernaute, Rosario" sort="Sanchez Pernaute, Rosario" uniqKey="Sanchez Pernaute R" first="Rosario" last="Sanchez-Pernaute">Rosario Sanchez-Pernaute</name><affiliation><nlm:affiliation>Laboratory of Stem Cells and Neural Repair and Cytometry and Advanced Optical Microscopy Facility, Inbiomed, San Sebastian, Spain; STEMCELL Technologies, Inc., Vancouver, British Columbia, Canada; Division of Gene Regulation, Netherlands Cancer Institute, Amsterdam, The Netherlands; Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden; Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA rpernaute@inbiomed.org.</nlm:affiliation><wicri:noCountry code="subField">USA rpernaute@inbiomed.org.</wicri:noCountry></affiliation></author></analytic><series><title level="j">Stem cells translational medicine</title><idno type="ISSN">2157-6564</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Blotting, Western</term><term>Cell Separation (methods)</term><term>Dopaminergic Neurons (cytology)</term><term>Flow Cytometry</term><term>Fluorescent Antibody Technique</term><term>Genetic Vectors</term><term>Green Fluorescent Proteins (genetics)</term><term>Hepatocyte Nuclear Factor 3-beta (genetics)</term><term>Humans</term><term>Lentivirus</term><term>Microscopy, Confocal</term><term>Neural Stem Cells (cytology)</term><term>Pluripotent Stem Cells (cytology)</term><term>Promoter Regions, Genetic</term><term>Real-Time Polymerase Chain Reaction</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Transduction, Genetic</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Hepatocyte Nuclear Factor 3-beta</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Dopaminergic Neurons</term><term>Neural Stem Cells</term><term>Pluripotent Stem Cells</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Cell Separation</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Blotting, Western</term><term>Flow Cytometry</term><term>Fluorescent Antibody Technique</term><term>Genetic Vectors</term><term>Humans</term><term>Lentivirus</term><term>Microscopy, Confocal</term><term>Promoter Regions, Genetic</term><term>Real-Time Polymerase Chain Reaction</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Transduction, Genetic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Human embryonic and induced pluripotent stem cells are potential cell sources for regenerative approaches in Parkinson disease. Inductive differentiation protocols can generate midbrain dopamine neurons but result in heterogeneous cell mixtures. Therefore, selection strategies are necessary to obtain uniform dopamine cell populations. Here, we developed a selection approach using lentivirus vectors to express green fluorescent protein under the promoter region of FOXA2, a transcription factor that is expressed in the floor plate domain that gives rise to dopamine neurons during embryogenesis. We first validated the specificity of the vectors in human cell lines against a promoterless construct. We then selected FOXA2-positive neural progenitors from several human pluripotent stem cell lines, which demonstrated a gene expression profile typical for the ventral domain of the midbrain and floor plate, but failed to enrich for dopamine neurons. To investigate whether this was due to the selection approach, we overexpressed FOXA2 in neural progenitors derived from human pluripotent stem cell lines. FOXA2 forced expression resulted in an increased expression of floor plate but not mature neuronal markers. Furthermore, selection of the FOXA2 overexpressing fraction also failed to enrich for dopamine neurons. Collectively, our results suggest that FOXA2 is not sufficient to induce a dopaminergic fate in this system. On the other hand, our study demonstrates that a combined approach of promoter activation and lentivirus vector technology can be used as a versatile tool for the selection of a defined cell population from a variety of human pluripotent stem cell lines.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Massachusetts</li></region></list><tree><noCountry><name sortKey="Sanchez Pernaute, Rosario" sort="Sanchez Pernaute, Rosario" uniqKey="Sanchez Pernaute R" first="Rosario" last="Sanchez-Pernaute">Rosario Sanchez-Pernaute</name></noCountry><country name="États-Unis"><region name="Massachusetts"><name sortKey="Aguila, Julio Cesar" sort="Aguila, Julio Cesar" uniqKey="Aguila J" first="Julio Cesar" last="Aguila">Julio Cesar Aguila</name></region><name sortKey="Aduriz, Ariane" sort="Aduriz, Ariane" uniqKey="Aduriz A" first="Ariane" last="Aduriz">Ariane Aduriz</name><name sortKey="Blak, Alexandra" sort="Blak, Alexandra" uniqKey="Blak A" first="Alexandra" last="Blak">Alexandra Blak</name><name sortKey="Gayosso, Mayela" sort="Gayosso, Mayela" uniqKey="Gayosso M" first="Mayela" last="Gayosso">Mayela Gayosso</name><name sortKey="Hedlund, Eva" sort="Hedlund, Eva" uniqKey="Hedlund E" first="Eva" last="Hedlund">Eva Hedlund</name><name sortKey="Lopez De Maturana, Rakel" sort="Lopez De Maturana, Rakel" uniqKey="Lopez De Maturana R" first="Rakel" last="Lopez De Maturana">Rakel Lopez De Maturana</name><name sortKey="Lopez Mato, Maria Paz" sort="Lopez Mato, Maria Paz" uniqKey="Lopez Mato M" first="Maria Paz" last="Lopez Mato">Maria Paz Lopez Mato</name><name sortKey="Sonntag, Kai Christian" sort="Sonntag, Kai Christian" uniqKey="Sonntag K" first="Kai-Christian" last="Sonntag">Kai-Christian Sonntag</name><name sortKey="Sousa, Amaia" sort="Sousa, Amaia" uniqKey="Sousa A" first="Amaia" last="Sousa">Amaia Sousa</name><name sortKey="Van Arensbergen, Joris" sort="Van Arensbergen, Joris" uniqKey="Van Arensbergen J" first="Joris" last="Van Arensbergen">Joris Van Arensbergen</name><name sortKey="Vazquez, Nerea" sort="Vazquez, Nerea" uniqKey="Vazquez N" first="Nerea" last="Vázquez">Nerea Vázquez</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Canada/explor/ParkinsonCanadaV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000934 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000934 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Canada
|area= ParkinsonCanadaV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:25024431
|texte= Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for Dopamine Neurons From Human Pluripotent Stem Cells.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:25024431" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a ParkinsonCanadaV1
| This area was generated with Dilib version V0.6.29. |  |