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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.
MACS Handbook:
Hematopoietic stem cells (human)
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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.
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Related videos:
How to enrich CD34+ cells from blood products with CliniMACS Technology?
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.
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