We isolated and enriched CD34+ cells from 2 large UCB units. A post enrichment count was 7x 10^6 million cells, we incubated overnight and this morning there are only 1 x 10^6. We have seen a small drop here or there, but never this significant.
Any ideas?
Hi Amanda, Thats a huge change in cell count. The possible answer could be cell death/apoptosis. It is not clear in which media you incubated them. If its serum free medium, then you need to supplement with at least suboptimal concentration of growth factors especially TPO and SCF. This reduce the cell death. But if you are using complete medium (10-20% serum), then you may see lose some cells because of attachment to the culture dish.
CD34+ cells from UCB are keeping a live for 24-36 hrs. It can divide as well. After that time it goes into death phase and/or apoptosis.
http://www.ncbi.nlm.nih.gov/pubmed/25473447
Thanks for the responses. We culture in STEMspan and Cytokine cocktails to reduce the differentiation. We cannot add serum to the mix as we are specifically looking at the progenitor line.
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