title: Characterization of P19 cells during retinoic acid induced differentiation.
abstract: The aim of our study was to characterize mouse embryonal carcinoma (EC) cells P19 in different stages of retinoic acid induced neurodifferentiation by two methods, immunocytochemistry and RT qPCR. The characterization of the cells is crucial before any transplantation into any model, e.g. in our case into the mouse brain with the aim to treat a neurodegenerative disease. Specific protein markers (MAP-2, OCT-4, FORSE-1) were detected by immunocytochemistry in the cell cultures. The mRNA expression levels of PAX-6, MASH-1, Brachyury, GATA-4 and AFP were determined by RT qPCR method. HPRT was used as a housekeeping gene. The degree of differentiation can be characterized by expression of analyzed genes. The presence of OCT-4 and FORSE-1 proteins in undifferentiated pluripotent cells and the presence of dendrite specific MAP-2 in neuroprogenitors was detected. The expression levels of PAX-6 and MASH-1 increased and expression of Brachyury decreased during the neurodifferentiation process. The expression levels of GATA-4 and AFP were the highest after induction of differentiation with retinoic acid. Detailed characterization of cells before transplantation experiments can contribute to better understanding of their effect.
authors.affiliation: Charles University in Prague, Faculty of Medicine in Plzeň, Department of Medical Chemistry and Biochemistry, Plzeň, Czech Republic. vaclav.babuska@lfp.cuni.cz
authors.forename: V
authors.initials: V
authors.lastname: Babuška
authors.forename: V
authors.initials: V
authors.lastname: Kulda
authors.forename: Z
authors.initials: Z
authors.lastname: Houdek
authors.forename: M
authors.initials: M
authors.lastname: Pešta
authors.forename: J
authors.initials: J
authors.lastname: Cendelín
authors.forename: N
authors.initials: N
authors.lastname: Zech
authors.forename: J
authors.initials: J
authors.lastname: Pacherník
authors.forename: F
authors.initials: F
authors.lastname: Vožeh
authors.forename: P
authors.initials: P
authors.lastname: Uher
authors.forename: M
authors.initials: M
authors.lastname: Králíčková
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics*; D045744:Cell Line, Tumor; D054278:Embryonal Carcinoma Stem Cells / Q000502:physiology; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D000954:Antigens, Surface; D051792:Basic Helix-Loop-Helix Proteins; D005136:Eye Proteins; D018398:Homeodomain Proteins; D008869:Microtubule-Associated Proteins; D050814:Octamer Transcription Factor-3; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins; D014212:Tretinoin; C065500:ASCL1 protein, human; C000605730:PAX6 protein, human; C000605732:Pax6 protein, mouse; C494174:Pou5f1 protein, mouse; C092260:antigen FORSE-1