Large-scale ex vivo expansion and characterization of natural killer cells for clinical applications
| Autor: | Susann Szmania, Juan F. Vera, Dario Campana, Natalia Lapteva, Elaine Coustan-Smith, April G. Durett, Amberly Moreno-Bost, Minhtran C. Ngo, Jiali Sun, Leslie L. Huye, Adrian P. Gee, Jun Ando, Lisa Rollins, Frits Vanrhee, Cliona M. Rooney, Jian Fang, Zhuyong Mei, Tarun K. Garg, Varada Dandekar, Kimberley Jackson |
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| Rok vydání: | 2012 |
| Předmět: |
Cancer Research
Cell Survival T-Lymphocytes medicine.medical_treatment Immunology Cell Culture Techniques Biology Lymphocyte Activation Immunotherapy Adoptive Article Flow cytometry medicine Humans Immunology and Allergy Ex vivo expansion Genetics (clinical) Cryopreservation Interleukin-15 Transplantation medicine.diagnostic_test Histocompatibility Antigens Class I Cell Biology Immunotherapy Flow Cytometry Killer Cells Natural 4-1BB Ligand Oncology Cell culture Interleukin 15 Blood Component Removal Cancer research Lymphocyte activation K562 Cells K562 cells Cell based |
| Zdroj: | Cytotherapy. 14:1131-1143 |
| ISSN: | 1465-3249 |
| Popis: | Interest in natural killer (NK) cell-based immunotherapy has resurged since new protocols for the purification and expansion of large numbers of clinical-grade cells have become available.We have successfully adapted a previously described NK expansion method that uses K562 cells expressing interleukin (IL)-15 and 4-1 BB Ligand (BBL) (K562-mb15-41BBL) to grow NK cells in novel gas-permeable static cell culture flasks (G-Rex).Using this system we produced up to 19 × 10(9) functional NK cells from unseparated apheresis products, starting with 15 × 10(7) CD3(-) CD56 (+) NK cells, within 8-10 days of culture. The G-Rex yielded a higher fold expansion of NK cells than conventional gas-permeable bags and required no cell manipulation or feeding during the culture period. We also showed that K562-mb15-41BBL cells up-regulated surface HLA class I antigen expression upon stimulation with the supernatants from NK cultures and stimulated alloreactive CD8 (+) T cells within the NK cultures. However, these CD3 (+) T cells could be removed successfully using the CliniMACS system. We describe our optimized NK cell cryopreservation method and show that the NK cells are viable and functional even after 12 months of cryopreservation.We have successfully developed a static culture protocol for large-scale expansion of NK cells in the gas permeable G-Rex system under good manufacturing practice (GMP) conditions. This strategy is currently being used to produce NK cells for cancer immunotherapy. |
| Databáze: | OpenAIRE |
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