A fluorescent reporter gene as a marker for ventricular specification in ES-derived cardiac cells

Autor: Meyer, M., Jaconi, Marisa, Landopoulou, Angela, Fort, Philippe, Puceat, Michel, Meyer, Nathalie
Přispěvatelé: Unité de Nutrition Humaine (UNH), Institut National de la Recherche Agronomique (INRA)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Clermont Université, Université de Genève = University of Geneva (UNIGE), Molecular Diagnostics Laboratory, National Center for Scientific Research 'Demokritos' (NCSR)-IRRP, Centre de recherches de biochimie macromoléculaire (CRBM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-IFR122-Centre National de la Recherche Scientifique (CNRS), Université d'Auvergne - Clermont-Ferrand I (UdA)-Clermont Université-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS)-IFR122-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), Equipe avenir HUMAN EMBRYONIC STEM CELLS AS A MODEL TO DELINEATE THE EARLY CARDIAC TRANSCRIPTIONAL NETWORKS: TOWARD A BETTER UNDERSTANDING OF CARDIAC CONGENITAL AND GENETIC DISEASES., Institut National de la Santé et de la Recherche Médicale (INSERM)
Jazyk: angličtina
Rok vydání: 2000
Předmět:
Myosin Light Chains/genetics
Time Factors
Cell division
Cellular differentiation
Embryoid bodies
Ryanodine Receptor Calcium Release Channel/analysis
RNA
Messenger/analysis/genetics
Embryoid body
[SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC]
ddc:616.07
[SDV.BID.SPT]Life Sciences [q-bio]/Biodiversity/Systematics
Phylogenetics and taxonomy
Biochemistry
Heart Ventricles/ cytology/ embryology/metabolism
Green fluorescent protein
MESH: Ryanodine Receptor Calcium Release Channel
Mice
Structural Biology
Genes
Reporter
embryonic stem-derived cardiac cellsventricular myosin light chain 2 promoterembryoid bodies
MESH: Reverse Transcriptase Polymerase Chain Reaction
MESH: Gene Expression Regulation
Developmental
Ventricular Function
MESH: Animals
Promoter Regions
Genetic
MESH: Cardiac Myosins
Ventricular myosin light chain 2 promoter
MESH: Organ Specificity
0303 health sciences
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells
030302 biochemistry & molecular biology
Luminescent Proteins/ genetics/metabolism
Gene Expression Regulation
Developmental
Cell Differentiation
Cell cycle
MESH: Myosin Light Chains
Organ Specificity
MESH: Cell Division
MESH: Luminescent Proteins
Cell Division
MESH: Cell Differentiation
Myosin Light Chains
MESH: Myocardium
Heart Ventricles
Biophysics
Color
[SDV.CAN]Life Sciences [q-bio]/Cancer
MESH: Stem Cells
Biology
Transfection
Fluorescence
Cell Line
03 medical and health sciences
Genes
Reporter/ genetics
Cell Clone
MESH: Promoter Regions
Genetic
MESH: Ventricular Function
Genetics
Animals
RNA
Messenger
Molecular Biology
MESH: Mice
Stem Cells/ cytology/metabolism
030304 developmental biology
MESH: RNA
Messenger
MESH: Color
Myocardium
MESH: Transfection
MESH: Time Factors
MESH: Fluorescence
MESH: Genes
Reporter
Ryanodine Receptor Calcium Release Channel
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
Cell Biology
Myocardium/ cytology/metabolism
Molecular biology
Embryonic stem cell
Promoter Regions
Genetic/genetics
MESH: Cell Line
Luminescent Proteins
[SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis
Cell culture
MESH: Heart Ventricles
Embryonic stem-derived cardiac cells
Cardiac Myosins
Zdroj: FEBS Letters
FEBS Letters, 2000, 478 (1-2), pp.151-158. ⟨10.1016/s0014-5793(00)01839-1⟩
FEBS Letters, Wiley, 2000, 478 (1-2), pp.151-158. ⟨10.1016/s0014-5793(00)01839-1⟩
FEBS Letters, Vol. 478, No 1-2 (2000) pp. 151-158
ISSN: 0014-5793
1873-3468
DOI: 10.1016/s0014-5793(00)01839-1⟩
Popis: International audience; We have established a CGR8 embryonic stem (ES) cell clone (MLC2ECFP) which expresses the enhanced cyan variant of Aequorea victoria green fluorescent protein (ECFP) under the transcriptional control of the ventricular myosin light chain 2 (MLC2v) promoter. Using epifluorescence imaging of vital embryoid bodies (EB) and reverse transcription-polymerase chain reaction (RT-PCR), we found that the MLC2v promoter is switched on as early as day 7 and is accompanied by formation of cell clusters featuring a bright ECFP blue fluorescence. The fluorescent areas within the EBs were all beating on day 8. MLC2ECFP ES cells showed the same time course of cardiac differentiation as mock ES cells as assessed by RT-PCR of genes encoding cardiac-specific transcription factors and contractile proteins. The MLC2v promoter conferred ventricular specificity to ECFP expression within the EB as revealed by MLC2v co-staining of ECFP fluorescent cells. MLC2ECFP-derived cardiac cells still undergo cell division on day 12 after isolation from EBs but withdraw from the cell cycle on day 16. This ES cell clone provides a powerful cell model to study the signalling roads of factors regulating cardiac cell proliferation and terminal differentiation with a view to using them for experimental cell therapy.
Databáze: OpenAIRE
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