[Comparative Study of the Two High-Efficient Strategies for Generation of Human Umbilical Cord Blood-derived Natural Killer Cells].

Objective: To explore the similarities and variations of biological phenotype and cytotoxicity of human umbilical cord blood natural killer cells (hUC- NK) after human umbilical cord blood-derived mononuclear cells (hUC-MNC) activated and expanded by two high-efficient strategies.

Methods: Umbilical cord blood mononuclear cells (MNC) from healthy donor were enriched by Ficoll-based density gradient centrifugation. Then, the phenotype, subpopulations, cell viability and cytotoxicity of NK cells derived from Miltenyi medium (denoted as M-NK) and X-VIVO 15 (denoted as X-NK) were compared using a "3IL" strategy.

Results: After a 14-day's culture, the contents of CD3CD56 NK cells were elevated from 4.25%±0.04% (d 0) to 71%±0.18% (M-NK) and 75.2%±1.1% (X-NK) respectively. Compared with X-NK group, the proportion of CD3CD4 T cells and CD3CD56 NKT cells in M-NK group decreased significantly. The percentages of CD16, NKG2D, NKp44, CD25 NK cells in X-NK group was higher than those in the M-NK group, while the total number of expanded NK cells in X-NK group was half of that in M-NK group. There were no significant differences between X-NK and M-NK groups in cell proliferation and cell cycle, except for the lower percentage of Annexin V+ apoptotic cells in M-NK group. Compared with X-NK group, the proportion of CD107a NK cells in M-NK group were higher under the same effector-target ratio (E∶T) (<0.05).

Conclusion: The two strategies were adequate for high-efficient generation of NK cells with high level of activation , however, there are differences in biological phenotypes and tumor cytotoxicity.