Generation of mouse-induced pluripotent stem cells by lentiviral transduction

Xiaodong Liu, Joseph Chen, Jaber Firas, Jacob M. Paynter, Christian M. Nefzger, Jose M. Polo

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

Abstract

Terminally differentiated somatic cells can be reprogrammed into an embryonic stem cell-like state by the forced expression of four transcription factors: Oct4, Klf4, Sox2, and c-Myc (OKSM). These so-called induced pluripotent stem (iPS) cells can give rise to any cell type of the body and thus have tremendous potential for many applications in research and regenerative medicine. Herein, we describe (1) a protocol for the generation of iPS cells from mouse embryonic fibroblasts (MEFs) using a doxycycline (Dox)-inducible lentiviral transduction system; (2) the derivation of clonal iPS cell lines; and (3) the characterization of the pluripotent potential of iPS cell lines using alkaline phosphatase staining, flow cytometry, and the teratoma formation assays.

Original languageEnglish
Title of host publicationMouse Cell Culture
Subtitle of host publicationMethods and Protocols
EditorsIvan Bertoncello
Place of PublicationNew York, NY
PublisherHumana Press
Chapter5
Pages63-76
Number of pages14
ISBN (Electronic)9781493990863
ISBN (Print)9781493990856
DOIs
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1940
ISSN (Print)1064-3745

Keywords

  • Fibroblasts
  • Lentiviral transduction
  • Mouse-induced pluripotent stem cells
  • OKSM
  • Reprogramming
  • Teratoma assay

Cite this

Liu, X., Chen, J., Firas, J., Paynter, J. M., Nefzger, C. M., & Polo, J. M. (2019). Generation of mouse-induced pluripotent stem cells by lentiviral transduction. In I. Bertoncello (Ed.), Mouse Cell Culture: Methods and Protocols (pp. 63-76). (Methods in Molecular Biology; Vol. 1940). Humana Press. https://doi.org/10.1007/978-1-4939-9086-3_5