Manufacturing of gene-engineered hematopoietic stem cells

Viral transduction of CD34⁺ cells is a promising approach for treating inherited disorders such as sickle cell disease, ß-thalassemia, or primary immunodeficiencies ^1,2,3. The CliniMACS Prodigy® Hematopoietic Stem Cell Engineering (HSCE) System allows for manufacturing of gene-engineered hematopoietic stem cells (HSCs) from human CD34⁺ cells.

The CliniMACS Prodigy Cell Manufacturing Platform automates the generation of gene-engineered HSCs by viral transduction in a functionally closed system. All steps are automated and manufacturing takes place in sterile and single-use tubing sets, ensuring highly reproducible and standardized manufacturing processes.

1. Preparation of CD34⁺ cells as cellular starting material 

The CliniMACS Prodigy HSCE System uses human CD34⁺ cells as cellular starting material. They can be easily enriched from mobilized leukapheresis with the CliniMACS Prodigy or the CliniMACS Plus Instrument. Enriched CD34⁺ cells are transferred to a cell bag, which needs to be connected to the CliniMACS Prodigy Tubing Set (TS) 520.

2. Transduction of HSCs

CD34⁺ cells are resuspended in HSC-Brew GMP Medium supplemented with MACS® GMP Recombinant Human Cytokines (SCT, TPO, Flt3-Ligand and IL-3) and transduced with lentiviral vector. The transduction step can be repeated, if necessary, to increase transduction efficiency.

3. Harvest of gene-engineered HSCs

After transduction, the final cell product containing gene-engineered HSCs is washed twice and harvested in 100 mL buffer of your choice. The gene-engineered HSCs are then ready for further processing, e.g., cryopreservation, or functional testing.
 

References

1. Lebensburger, J. and Persons D.A. (2008) Curr. Opin. Drug Discov. Devel. 11: 225–232.
2. Papanikolaou E. and Anagnou N.P. (2010) Curr. Gene Ther. 10: 404–412.
3. de Dreuzy E. et al. (2016) Biomed. J. 39: 24–38.