title: Transplantation of embryonic neuroectodermal progenitor cells into the site of a photochemical lesion: immunohistochemical and electrophysiological analysis.
abstract: GFP labeled/NE-4C neural progenitor cells cloned from primary neuroectodermal cultures of p53- mouse embryos give rise to neurons when exposed to retinoic acid in vitro. To study their survival and differentiation in vivo, cells were transplanted into the cortex of 6-week-old rats, 1 week after the induction of a photochemical lesion or into noninjured cortex. The electrophysiological properties of GFP/NE-4C cells were studied in vitro (8-10 days after differentiation induction) and 4 weeks after transplantation using the whole-cell patch-clamp technique, and immunohistochemical analyses were carried out. After transplantation into a photochemical lesion, a large number of cells survived, some of which expressed the astrocytic marker GFAP. GFP/GFAP-positive cells, with an average resting membrane potential (Vrest) of -71.9 mV, displayed passive time- and voltage-independent K+ currents and, additionally, voltage-dependent A-type K+ currents (KA) and/or delayed outwardly rectifying K+ currents (KDR). Numerous GFP-positive cells expressed NeuN, betaIII-tubulin, or 68 kD neurofilaments. GFP/betaIII-tubulin-positive cells, with an average Vrest of -61.6 mV, were characterized by the expression of KA and KDR currents and tetrodotoxin-sensitive Na+ currents. GFP/NE-4C cells also gave rise to oligodendrocytes, based on the detection of oligodendrocyte-specific markers. Our results indicate that GFP/NE-4C neural progenitors transplanted into the site of a photochemical lesion give rise to neurons and astrocytes with membrane properties comparable to those transplanted into noninjured cortex. Therefore, GFP/NE-4C cells provide a suitable model for studying neuro- and gliogenesis in vivo. Further, our results suggest that embryonic neuroectodermal progenitor cells may hold considerable promise for the repair of ischemic brain lesions.
authors.affiliation: Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic. anderova@biomed.cas.cz
authors.forename: Miroslava
authors.initials: M
authors.lastname: Anderová
authors.forename: Sárka
authors.initials: S
authors.lastname: Kubinová
authors.forename: Marti
authors.initials: M
authors.lastname: Jelitai
authors.forename: Helena
authors.initials: H
authors.lastname: Neprasová
authors.forename: Katerina
authors.initials: K
authors.lastname: Glogarová
authors.forename: Iva
authors.initials: I
authors.lastname: Prajerová
authors.forename: Lucie
authors.initials: L
authors.lastname: Urdzíková
authors.forename: Alexandr
authors.initials: A
authors.lastname: Chvátal
authors.forename: Eva
authors.initials: E
authors.lastname: Syková
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001253:Astrocytes / Q000502:physiology; D002545:Brain Ischemia / Q000473:pathology / Q000628:therapy*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002540:Cerebral Cortex / Q000473:pathology / Q000502:physiology / Q000601:surgery; D003714:Denervation / Q000379:methods; D004195:Disease Models, Animal; D004475:Ectoderm / Q000166:cytology; D006085:Graft Survival; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D008564:Membrane Potentials; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D009836:Oligodendroglia / Q000502:physiology; D018408:Patch-Clamp Techniques; D017319:Photosensitizing Agents; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D017319:Photosensitizing Agents; D049452:Green Fluorescent Proteins; D014212:Tretinoin