Expansion of Human CD34+ Hematopoietic Stem Cells
Human CD34+ HSPCs were cultured as previously described
20 (link),21 (link). CD34+ HSPCs were purchased from AllCells and had been isolated from G-CSF-mobilized peripheral blood from healthy donors. CD34+ HSPCs were cultured at 2.5 × 10
5–5 × 10
5 cells/mL in
StemSpan™-AOF (Stemcell) supplemented with stem cell factor (SCF) (100 ng/mL), thrombopoietin (TPO) (100 ng/mL),
FLT3–ligand (100 ng/mL),
IL-6 (100 ng/mL) (all Peprotech) and
UM171 (35 nM) (Selleckchem). Cells were cultured at 37 °C, 5% CO
2, and 5% O
2.
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Perez-Bermejo J.A., Efagene O., Matern W.M., Holden J.K., Kabir S., Chew G.M., Andreoletti G., Catton E., Ennis C.L., Garcia A., Gerstenberg T.L., Hill K.A., Jain A., Krassovsky K., Lalisan C.D., Lord D., Quejarro B.J., Sales-Lee J., Shah M., Silva B.J., Skowronski J., Strukov Y.G., Thomas J., Veraz M., Vijay T., Wallace K.A., Yuan Y., Grogan J.L., Wienert B., Lahiri P., Treusch S., Dever D.P., Soros V.B., Partridge J.R, & Seim K.L. (2024). Functional screening in human HSPCs identifies optimized protein-based enhancers of Homology Directed Repair. Nature Communications, 15, 2625.
