High-Efficiency Lentiviral Transduction of Human CD34+ Cells in High-Density Culture with Poloxamer and Prostaglandin E2

Molecular Therapy. Methods & Clinical Development
Naoya UchidaJohn F Tisdale

Abstract

Hematopoietic stem cell (HSC) gene therapy is curative for various hereditary diseases; however, high-efficiency transduction in HSCs remains crucial to improve the prospects for hemoglobinopathies. We previously optimized lentiviral transduction in human CD34+ cells with serum-free medium containing minimal cytokines, allowing efficient transduction (∼50%) and robust xenograft engraftment. In this study, we further improved lentiviral transduction in human CD34+ cells. High-density culture conditions (4e6/mL) resulted in ∼5-fold more efficient transduction in CD34+ cells (p < 0.01) compared with standard cell density (1e5/mL). After co-culturing vector-exposed CD34+ cells with non-transduced CD34+ cells, high-density culture conditions enhanced lentiviral gene marking in the non-transduced population (p < 0.01) compared with low-density conditions, suggesting that increasing cell-to-cell contact allows more efficient transduction. Two adjuvants, poloxamer 407 (100 μg/mL) and prostaglandin E2 (10 μM), were added to high-density CD34+ cells, resulting in ∼4-fold more efficient transduction (p < 0.01) without significant toxicity compared with no adjuvant control. In summary, we developed a highly efficient lentiviral transductio...Continue Reading

Citations

Oct 4, 2019·Nature Communications·Naoya UchidaJohn F Tisdale
Feb 2, 2021·Frontiers in Cardiovascular Medicine·Jose Manuel Condor CapchaLina A Shehadeh
Apr 6, 2021·Molecular Therapy. Methods & Clinical Development·Naoya UchidaJohn F Tisdale
Apr 27, 2021·Molecular Therapy. Methods & Clinical Development·Vania Lo PrestiNiek P van Til
Aug 23, 2019·International Journal of Biological Macromolecules·Jonathan Tersur OrasughDipankar Chattopadhyay
Apr 18, 2020·Molecular Therapy. Methods & Clinical Development·Christian BrendelDavid A Williams
May 15, 2020·Molecular Therapy. Methods & Clinical Development·Selami DemirciNaoya Uchida
Jul 8, 2021·Blood·Allistair A Abraham, John F Tisdale

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Methods Mentioned

BETA
xenograft
flow
PCR
flow cytometry

Software Mentioned

QuantStudio
JMP

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