title: Clonal human fetal ventral mesencephalic dopaminergic neuron precursors for cell therapy research.
abstract: A major challenge for further development of drug screening procedures, cell replacement therapies and developmental studies is the identification of expandable human stem cells able to generate the cell types needed. We have previously reported the generation of an immortalized polyclonal neural stem cell (NSC) line derived from the human fetal ventral mesencephalon (hVM1). This line has been biochemically, genetically, immunocytochemically and electrophysiologically characterized to document its usefulness as a model system for the generation of A9 dopaminergic neurons (DAn). Long-term in vivo transplantation studies in parkinsonian rats showed that the grafts do not mature evenly. We reasoned that diverse clones in the hVM1 line might have different abilities to differentiate. In the present study, we have analyzed 9 hVM1 clones selected on the basis of their TH generation potential and, based on the number of v-myc copies, v-myc down-regulation after in vitro differentiation, in vivo cell cycle exit, TH⁺ neuron generation and expression of a neuronal mature marker (hNSE), we selected two clones for further in vivo PD cell replacement studies. The conclusion is that homogeneity and clonality of characterized NSCs allow transplantation of cells with controlled properties, which should help in the design of long-term in vivo experiments.
authors.affiliation: Department of Molecular Biology-U.A.M. and Center of Molecular Biology "Severo Ochoa"-U.A.M.-C.S.I.C. Nicolás Cabrera, 1 Universidad Autónoma de Madrid, Campus Cantoblanco, Madrid, Spain. tramos@cbm.uam.es
authors.forename: Tania
authors.initials: T
authors.lastname: Ramos-Moreno
authors.forename: Javier G
authors.initials: JG
authors.lastname: Lendínez
authors.forename: María José
authors.initials: MJ
authors.lastname: Pino-Barrio
authors.forename: Araceli
authors.initials: A
authors.lastname: Del Arco
authors.forename: Alberto
authors.initials: A
authors.lastname: Martínez-Serrano
mesh_terms: D000818:Animals; D059447:Cell Cycle Checkpoints; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D059290:Dopaminergic Neurons / Q000502:physiology* / Q000637:transplantation; D015536:Down-Regulation; D005260:Female; D018628:Gene Dosage; D015870:Gene Expression; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D019394:Ki-67 Antigen / Q000235:genetics / Q000378:metabolism; D008636:Mesencephalon; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D058953:Neural Stem Cells / Q000502:physiology* / Q000637:transplantation; D016274:Oncogene Protein p55(v-myc) / Q000235:genetics / Q000378:metabolism; D010302:Parkinson Disease, Secondary / Q000628:therapy*; D051381:Rats; D017207:Rats, Sprague-Dawley; D057905:Stem Cell Research
chemical_list: D007381:Intermediate Filament Proteins; D019394:Ki-67 Antigen; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D016274:Oncogene Protein p55(v-myc)