High-efficiency lentiviral transduction of primary human CD34+ hematopoietic cells with low-dose viral inocula

Sang Won Park, Chul Woong Pyo, Sang-Yun Choi

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Lentivirus-based vectors have the potential to transduce non-dividing primary stem cells. However, primary cells tend to be less susceptible to manipulation and require a high concentration of virus inoculum. Furthermore, increasing the concentration of the lentivirus inoculum may raise safety risks. Therefore, to develop a technique that allows high transduction efficiency at low multiplicities of infection (MOIs), we optimized a lentivirus-based system for cell lines and primary cells by determining the best condition using various parameters. When progenitor cell assays were conducted using human CD34+ bone marrow and mononuclear cells, the transduction condition yielded a great number of eGFP+ colonies with lower-dose viral inocula compared to that of current lentivirus-based transduction technologies. In conclusion, this system is anticipated to produce stable expression of a gene introduced into primary cells for preclinical studies with lower safety risks.

Original languageEnglish
Pages (from-to)281-288
Number of pages8
JournalBiotechnology Letters
Volume37
Issue number2
DOIs
Publication statusPublished - 2014

Keywords

  • CD34 bone marrow
  • Gene therapy
  • Hematopoietic cells
  • Progenitor cell assay
  • Pseudotyped lentiviral vector
  • Transduction efficiency
  • Viral inoculum

ASJC Scopus subject areas

  • Biotechnology

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