Early neuronal and glial determination from mouse E10.5 telencephalon embryonic stem cells: an in vitro study.
We report here a novel in vitro model for differentiating neuronal and glial cells from mouse embryonic day 10 telencephalon stem cells. At this developmental stage, the telencephalon consists of a single layer of neuroepithelial stem cells. We used various markers of proliferation and differentiation (Ki-67, nestin, BrdU, Tuj-1 and GFAP) to follow proliferative progenitors and to identify neuronal and glial derivatives. Neuronal derivatives were obtained from nestin+ progenitors. GFAP+ astrocytic derivatives were detected after only 72 h of culture. Both neuronal and glial derivatives were generated close to nestin-positive aggregates. In addition, we were able to manipulate neuronal determination of telencephalon stem cells by gene transient transfection as demonstrated by RP42 gene overexpression. These observations suggest that this in vitro model is of potential use for studying early steps in neuronal or glial determination from embryonic stem cells, an issue of key importance for adult brain cell therapy approaches.
Khelfaoui
Malik
M
Neurogénétique/INSERM E9935, Hôpital Robert Debré, 48 Boulevard Sérurier, 75019 Paris, France.
Guimiot
Fabien
F
Simonneau
Michel
M
D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D001973:Bromodeoxyuridine; D002454:Cell Differentiation / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics; D005260:Female; D005333:Fetus; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D007381:Intermediate Filament Proteins / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D019394:Ki-67 Antigen / Q000378:metabolism; D008164:Luminescent Proteins; D051379:Mice; D023421:Models, Animal; D009419:Nerve Tissue Proteins; D064231:Nestin; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy; D011506:Proteins / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013687:Telencephalon / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D014162:Transfection; D014404:Tubulin / Q000378:metabolism
C408631:DCUN1D1 protein, human; D005904:Glial Fibrillary Acidic Protein; D007202:Indicators and Reagents; D007381:Intermediate Filament Proteins; D047908:Intracellular Signaling Peptides and Proteins; D019394:Ki-67 Antigen; D008164:Luminescent Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D011506:Proteins; D014404:Tubulin; D049452:Green Fluorescent Proteins; D001973:Bromodeoxyuridine