Can you please provide me a protocol for the expansion of CD34+ Progenitor Cells in PromoCell Expansion Medium XF?

Short protocol:

• Thaw the cells (C-12921) for 2 min in a 37°C waterbath. Dilute in 9 ml of complete HPC Expansion Medium XF (+ Cytokine Mix E) and count the cells
• Spin down for 10 min at 240xg; aspirate the supernatant; resuspend the pellet at 20;000 cells/ml HPC Expansion Medium XF
• Plate in an appropriate suspension culture vessel and incubate the culture for 2-3 days at 37°C and 5% CO₂
• Then double the media volume by adding fresh complete medium; e.g.; 4 ml  suspension culture + 4 ml fresh medium (= 8 ml)
• Incubate the cells for an additional 10-12 days by performing a partial medium change every 2-3 days Example partial medium change: For a culture volume of 8 ml; spin down the cells; aspirate and discard 4 ml of the supernatant; resuspend the cells and add 12 ml of fresh complete medium (= 16 ml).

In combination with the Cytokine Mix E; the HPC Expansion Medium XF typically promotes a 300-1;000 fold expansion of the total cell population. After 2 weeks of expansion about 20-30% of the population express CD34⁺; indicating a 50-200 fold expansion of CD34⁺ progenitor cells.