Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment

Aaron D Schimmer; Rashim Pal Singh; Ayesh K Seneviratne; Geethu E Thomas; Neil MacLean; Rose Hurren

doi:10.1016/j.xpro.2020.100163

Transduction of Primary AML Cells with Lentiviral Vector for In Vitro Study or In Vivo Engraftment

STAR Protoc. 2020 Nov 25;1(3):100163. doi: 10.1016/j.xpro.2020.100163. eCollection 2020 Dec 18.

Authors

Aaron D Schimmer¹, Rashim Pal Singh¹, Ayesh K Seneviratne¹, Geethu E Thomas¹, Neil MacLean¹, Rose Hurren¹

Affiliation

¹ Princess Margaret Cancer Centre, University Health Network, Room 8-706, 101 College St., Toronto, ON M5G1L7, USA.

Abstract

We describe a method to silence genes in primary acute myeloid leukemia cells by transducing them with shRNA in lentiviral vectors. The transduction of primary non-adherent cells is particularly challenging. The protocol will aid in performing such experiments and is particularly helpful to prepare cells for in vivo engraftment studies. Use of a special medium supplemented with cytokines preserves the viability of the leukemic stem cells and their ability to engraft the marrow of immune-deficient mice. For complete details on the use and execution of this protocol, please refer to Singh et al. (2020).

Keywords: Cancer; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Genetic Vectors / metabolism*
Humans
Lentivirus / metabolism*
Leukemia, Myeloid, Acute / genetics*
Mice
Neoplasm Transplantation*
Transduction, Genetic*
Tumor Cells, Cultured

Grants and funding

CIHR/Canada