Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity

Cells

WinfriedS. Wels¹  Pranav Oberoi¹  Barbara Uherek¹  JoseFrancisco Villena Ossa¹  Kathrina Kamenjarin¹  Halvard Bönig² 

[1] Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, 60596 Frankfurt am Main, Germany;Goethe University, Institute for Transfusion Medicine and Immunohematology, and German Red Cross Blood Donation Service Baden-Württemberg - Hessen, 60528 Frankfurt am Main, Germany;

关键词: natural killer cells;    hematopoietic stem and progenitor cells;    feeder cells;    natural cytotoxicity;    adoptive cancer immunotherapy;   

DOI  :  10.3390/cells9040811

来源: DOAJ

【 摘 要 】

Obtaining sufficient numbers of functional natural killer (NK) cells is crucial for the success of NK-cell-based adoptive immunotherapies. While expansion from peripheral blood (PB) is the current method of choice, ex vivo generation of NK cells from hematopoietic stem and progenitor cells (HSCs) may constitute an attractive alternative. Thereby, HSCs mobilized into peripheral blood (PB-CD34⁺) represent a valuable starting material, but the rather poor and donor-dependent differentiation of isolated PB-CD34⁺ cells into NK cells observed in earlier studies still represents a major hurdle. Here, we report a refined approach based on ex vivo culture of PB-CD34⁺ cells with optimized cytokine cocktails that reliably generates functionally mature NK cells, as assessed by analyzing NK-cell-associated surface markers and cytotoxicity. To further enhance NK cell expansion, we generated K562 feeder cells co-expressing 4-1BB ligand and membrane-anchored IL-15 and IL-21. Co-culture of PB-derived NK cells and NK cells that were ex-vivo-differentiated from HSCs with these feeder cells dramatically improved NK cell expansion, and fully compensated for donor-to-donor variability observed during only cytokine-based propagation. Our findings suggest mobilized PB-CD34⁺ cells expanded and differentiated according to this two-step protocol as a promising source for the generation of allogeneic NK cells for adoptive cancer immunotherapy.

【 授权许可】

Unknown