Summary
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.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Ye, Z., Yu, X., Cheng, L. (2008). Lentiviral Gene Transduction of Mouse and Human Stem Cells. In: Bunting, K.D. (eds) Hematopoietic Stem Cell Protocols. Methods in Molecular Biology™, vol 430. Humana Press. https://doi.org/10.1007/978-1-59745-182-6_17
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DOI: https://doi.org/10.1007/978-1-59745-182-6_17
Publisher Name: Humana Press
Print ISBN: 978-1-58829-868-3
Online ISBN: 978-1-59745-182-6
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