A truncated SV40 large T antigen lacking the p53 binding domain overcomes p53-induced growth arrest and immortalizes primary mesencephalic cells

Cell and Tissue Research
Mary Ellen TruckenmillerW Freed

Abstract

As an alternative to primary fetal tissue, immortalized central nervous system (CNS)-derived cell lines are useful for in vitro CNS model systems and for gene manipulation with potential clinical use in neural transplantation. However, obtaining immortalized cells with a desired phenotype is unpredictable, because the molecular mechanisms of growth and differentiation of CNS cells are poorly understood. The SV40 large T antigen is commonly used to immortalize mammalian cells, but it interferes with multiple cell-cycle components, including p53, p300, and retinoblastoma protein, and usually produces cells with undifferentiated phenotypes. In order to increase the phenotypic repertoire of immortalized CNS cells and to address the molecular mechanisms underlying immortalization and differentiation, we constructed an expression vector containing a truncated SV40 large T gene that encodes only the amino-terminal 155 amino acids (T155), which lacks the p53-binding domain. Constructs were first transfected into a p53-temperature-sensitive cell line, T64-7B. Colonies expressing T155 proliferated at the growth-restrictive temperature. T155 was then transfected into primary cultures from embryonic day-14 rat mesencephalon. Two clonal cel...Continue Reading

Citations

Jan 13, 2006·Cell and Tissue Research·Joseph F SanchezWilliam J Freed
Apr 1, 2009·Neurotherapeutics : the Journal of the American Society for Experimental NeuroTherapeutics·Kerry Thompson
Dec 3, 2009·Viral Immunology·Paul M D'agostinoCarol Shoshkes Reiss
Jul 23, 1999·Annals of the New York Academy of Sciences·R JesnowskiM Löhr
Jul 27, 2001·Virchows Archiv : an International Journal of Pathology·M LöhrR Jesnowski
Aug 7, 2012·Journal of Cellular and Molecular Medicine·A Alwin Prem AnandV Kokila Vani
Apr 20, 2005·Brain Research. Molecular Brain Research·Jia ChenWilliam J Freed
Jan 5, 2005·Experimental Neurology·William J FreedMary Ellen Truckenmiller
Dec 25, 2002·The Tohoku Journal of Experimental Medicine·Masahiro SatohTooru Shimosegawa
May 25, 2010·International Journal for Parasitology·T QuackC G Grevelding

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