Springer Protocols: Abstract: Lentiviral Gene Transduction of Mouse and Human Stem Cells

Lentiviral Gene Transduction of Mouse and Human Stem Cells

By: Zhaohui Ye³, Xiaobing Yu³, Linzhao Cheng³

Abstract

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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.

Affiliation(s): (3) Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD

Book Title: Hematopoietic Stem Cell Protocols

Series: Methods in Molecular Biology | Volume: 430 | Pub. Date: Jan-01-2008 | Page Range: 243-253 | DOI: 10.1007/978-1-59745-182-6_17

Subject: Cell Biology

Key Words: Lentivirus - hematopoietic stem cells - hematopoietic progenitor cells - embryonic stem cells

References

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