Lentiviral gene transduction of mouse and human stem cells

Zhaohui Ye, Xiaobing Yu, Linzhao Cheng

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

This chapter describes the methods we use to transduce mouse and human hematopoietic stem cells (HSCs) and human embryonic stem cells (hESCs). We provide detailed protocols for producing high-titer lentiviral supernatants by transient transfection and for measuring viral titers. Methods to concentrate viral supernatants to achieve a higher titer are also described. The protocols given here have been used successfully to transduce engrafting mouse and human HSCs as well as progenitor cells. These cells maintained stable transgene expression after engraftment in mice and in vivo differentiation. Human ESCs can also be transduced with a high efficiency, and transgene is expressed stably after hematopoietic differentiation.

Original languageEnglish (US)
Title of host publicationHematopoietic Stem Cell Protocols
Pages243-253
Number of pages11
StatePublished - Aug 31 2007
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume430
ISSN (Print)1064-3745

Keywords

  • Embryonic stem cells
  • Hematopoietic progenitor cells
  • Hematopoietic stem cells
  • Lentivirus

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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