Protocol to produce high-titer retrovirus for transduction of mouse bone marrow cells

STAR Protoc. 2021 Apr 7;2(2):100459. doi: 10.1016/j.xpro.2021.100459. eCollection 2021 Jun 18.

Abstract

Regulating gene expression through retroviral infection has been widely used in mouse bone marrow transplantation (BMT) to test the capacity of self-renewal, as well as multi-lineage differentiation of hematopoietic stem and progenitor cells (HSPCs). However, it remains challenging to achieve high transduction efficiency in bone marrow cells as transduction of these cells subsequently leads to transplantation failure. Here, we present a modified protocol to overcome this issue, enabling reproducible and high-efficient retroviral transduction of HSPCs for BMT. For complete details on the use and execution of this protocol, please refer to Yang et al. (2019).

Keywords: Cell biology; Cell culture; Cell isolation; Model organisms; Stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allografts
  • Animals
  • Bone Marrow Transplantation
  • Gene Expression Regulation, Viral*
  • Hematopoietic Stem Cells / virology*
  • Mice
  • Retroviridae / genetics
  • Retroviridae / isolation & purification*
  • Retroviridae / metabolism*
  • Transduction, Genetic*