Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity
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) ma...
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MDPI AG
2020-03-01
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| Online Access: | https://www.mdpi.com/2073-4409/9/4/811 |
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| author | Pranav Oberoi Kathrina Kamenjarin Jose Francisco Villena Ossa Barbara Uherek Halvard Bönig Winfried S. Wels |
| author_facet | Pranav Oberoi Kathrina Kamenjarin Jose Francisco Villena Ossa Barbara Uherek Halvard Bönig Winfried S. Wels |
| author_sort | Pranav Oberoi |
| collection | DOAJ |
| description | 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<sup>+</sup>) represent a valuable starting material, but the rather poor and donor-dependent differentiation of isolated PB-CD34<sup>+</sup> 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<sup>+</sup> 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<sup>+</sup> 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. |
| first_indexed | 2024-03-11T10:12:13Z |
| format | Article |
| id | doaj.art-d3b05786b3074976aaebf85440fae1b3 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T10:12:13Z |
| publishDate | 2020-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-d3b05786b3074976aaebf85440fae1b32023-11-16T14:29:57ZengMDPI AGCells2073-44092020-03-019481110.3390/cells9040811Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor ActivityPranav Oberoi0Kathrina Kamenjarin1Jose Francisco Villena Ossa2Barbara Uherek3Halvard Bönig4Winfried S. Wels5Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, 60596 Frankfurt am Main, GermanyGeorg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, 60596 Frankfurt am Main, GermanyGeorg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, 60596 Frankfurt am Main, GermanyGeorg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, 60596 Frankfurt am Main, GermanyGoethe University, Institute for Transfusion Medicine and Immunohematology, and German Red Cross Blood Donation Service Baden-Württemberg - Hessen, 60528 Frankfurt am Main, GermanyGeorg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, 60596 Frankfurt am Main, GermanyObtaining 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<sup>+</sup>) represent a valuable starting material, but the rather poor and donor-dependent differentiation of isolated PB-CD34<sup>+</sup> 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<sup>+</sup> 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<sup>+</sup> 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.https://www.mdpi.com/2073-4409/9/4/811natural killer cellshematopoietic stem and progenitor cellsfeeder cellsnatural cytotoxicityadoptive cancer immunotherapy |
| spellingShingle | Pranav Oberoi Kathrina Kamenjarin Jose Francisco Villena Ossa Barbara Uherek Halvard Bönig Winfried S. Wels Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity Cells natural killer cells hematopoietic stem and progenitor cells feeder cells natural cytotoxicity adoptive cancer immunotherapy |
| title | Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity |
| title_full | Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity |
| title_fullStr | Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity |
| title_full_unstemmed | Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity |
| title_short | Directed Differentiation of Mobilized Hematopoietic Stem and Progenitor Cells into Functional NK Cells with Enhanced Antitumor Activity |
| title_sort | directed differentiation of mobilized hematopoietic stem and progenitor cells into functional nk cells with enhanced antitumor activity |
| topic | natural killer cells hematopoietic stem and progenitor cells feeder cells natural cytotoxicity adoptive cancer immunotherapy |
| url | https://www.mdpi.com/2073-4409/9/4/811 |
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