Optimization of SCF feeding regimen for ex vivo expansion of cord blood hematopoietic stem cells

Stem cell factor (SCF) plays important roles in ex vivo expansion of hematopoietic stem cells (HSCs). In this study, the effects of dose and feeding time of SCF on ex vivo expansion of CD34+ cells were investigated in serum-free medium supplemented with a cytokine cocktail composed of SCF, thrombopoietin (TPO) and flt3-ligand (FL). Among the four tested doses (0, 5, 50 and 500 ng/mL), a SCF dose of 50 ng/mL was demonstrated to be most favorable for ex vivo expansion of CD34+ cells, which resulted in 34.22 ± 10.80 and 8.89 ± 1.25 folds of expansion regarding total cells and CD34+ cells, respectively. Meanwhile, the specific growth rate of cells, the consumption rate of SCF and the percentage of CD34+c-kit+ cells during the 21-day culture process were analyzed. The results indicated that initial 4-day period was a critical stage for SCF functioning on CD34+ cells during ex vivo expansion. Based on this, a modified SCF feeding regimen was proposed, in which SCF (50 ng/mL) was only supplemented on day 0 in the cytokine cocktail and cells were then fed with TPO and FL till the end of culture. It was found that this SCF feeding regimen could expand CD34+ cells efficiently, thus providing a cost–effect expansion protocol for HSCs.

► The relationship between SCF dose in culture and the expansion of HSCs was studied.
► The initial 4 days were critical for SCF exerting function on HSCs ex vivo expansion.
► A new SCF feeding regimen in HSCs ex vivo culture was provided.
► This new feeding regimen could expand HSCs effectively.