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). New York, NY: Humana Press. https://doi.org/10.1007/978-1-4939-9086-3_5
Liu, Xiaodong ; Chen, Joseph ; Firas, Jaber ; Paynter, Jacob M. ; Nefzger, Christian M. ; Polo, Jose M. / Generation of mouse-induced pluripotent stem cells by lentiviral transduction. Mouse Cell Culture: Methods and Protocols. editor / Ivan Bertoncello. New York, NY : Humana Press, 2019. pp. 63-76 (Methods in Molecular Biology).
@inbook{0c1ce08d53474ea285cf0ef38777398c,
title = "Generation of mouse-induced pluripotent stem cells by lentiviral transduction",
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.",
keywords = "Fibroblasts, Lentiviral transduction, Mouse-induced pluripotent stem cells, OKSM, Reprogramming, Teratoma assay",
author = "Xiaodong Liu and Joseph Chen and Jaber Firas and Paynter, {Jacob M.} and Nefzger, {Christian M.} and Polo, {Jose M.}",
year = "2019",
doi = "10.1007/978-1-4939-9086-3_5",
language = "English",
isbn = "9781493990856",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "63--76",
editor = "Ivan Bertoncello",
booktitle = "Mouse Cell Culture",
address = "United States of America",

}

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

Generation of mouse-induced pluripotent stem cells by lentiviral transduction. / Liu, Xiaodong; Chen, Joseph; Firas, Jaber; Paynter, Jacob M.; Nefzger, Christian M.; Polo, Jose M.

Mouse Cell Culture: Methods and Protocols. ed. / Ivan Bertoncello. New York, NY : Humana Press, 2019. p. 63-76 (Methods in Molecular Biology; Vol. 1940).

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

TY - CHAP

T1 - Generation of mouse-induced pluripotent stem cells by lentiviral transduction

AU - Liu, Xiaodong

AU - Chen, Joseph

AU - Firas, Jaber

AU - Paynter, Jacob M.

AU - Nefzger, Christian M.

AU - Polo, Jose M.

PY - 2019

Y1 - 2019

N2 - 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.

AB - 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.

KW - Fibroblasts

KW - Lentiviral transduction

KW - Mouse-induced pluripotent stem cells

KW - OKSM

KW - Reprogramming

KW - Teratoma assay

UR - http://www.scopus.com/inward/record.url?scp=85061993651&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-9086-3_5

DO - 10.1007/978-1-4939-9086-3_5

M3 - Chapter (Book)

SN - 9781493990856

T3 - Methods in Molecular Biology

SP - 63

EP - 76

BT - Mouse Cell Culture

A2 - Bertoncello, Ivan

PB - Humana Press

CY - New York, NY

ER -

Liu X, Chen J, Firas J, Paynter JM, Nefzger CM, Polo JM. Generation of mouse-induced pluripotent stem cells by lentiviral transduction. In Bertoncello I, editor, Mouse Cell Culture: Methods and Protocols. New York, NY: Humana Press. 2019. p. 63-76. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9086-3_5