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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.
AuthorsV Babuška, V Kulda, Z Houdek, M Pešta, J Cendelín, N Zech, J Pacherník, F Vožeh, P Uher, M Králíčková
JournalPrague medical report (Prague Med Rep) Vol. 111 Issue 4 Pg. 289-99 ( 2010) ISSN: 1214-6994 [Print] Czech Republic
PMID21189168 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • ASCL1 protein, human
  • Antigens, Surface
  • Basic Helix-Loop-Helix Transcription Factors
  • Eye Proteins
  • Homeodomain Proteins
  • Microtubule-Associated Proteins
  • Octamer Transcription Factor-3
  • PAX6 Transcription Factor
  • PAX6 protein, human
  • Paired Box Transcription Factors
  • Pax6 protein, mouse
  • Pou5f1 protein, mouse
  • Repressor Proteins
  • antigen FORSE-1
  • Tretinoin
Topics
  • Animals
  • Antigens, Surface (genetics, metabolism)
  • Basic Helix-Loop-Helix Transcription Factors (genetics, metabolism)
  • Cell Differentiation (drug effects, genetics)
  • Cell Line, Tumor
  • Embryonal Carcinoma Stem Cells (physiology)
  • Eye Proteins (genetics, metabolism)
  • Gene Expression
  • Homeodomain Proteins (genetics, metabolism)
  • Immunohistochemistry
  • Mice
  • Microtubule-Associated Proteins (genetics, metabolism)
  • Octamer Transcription Factor-3 (genetics, metabolism)
  • PAX6 Transcription Factor
  • Paired Box Transcription Factors (genetics, metabolism)
  • Repressor Proteins (genetics, metabolism)
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tretinoin (pharmacology)
  • Tumor Cells, Cultured

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