Generation of p53-deficient induced pluripotent stem cells from mouse embryo fibroblasts

Abstract

Here we describe a method for generating induced pluripotent stem (iPS) cells from mouse embryonic fibroblasts (MEFs). Recombinant retroviruses carrying human transcription factors for Klf4, Oct3/4, Sox2, with or without c-Myc, are used to transduce early passage MEFs several times. Based on morphologic criteria, the resulting iPS colonies are picked manually at first, and then propagated and expanded by standard methods. iPS cells can then be differentiated into virtually any cell type or lineage, thus allowing for discoveries of new functions of p53 and mutant p53.