Streamlined production of genetically modified T cells with activation, transduction and expansion in closed-system G-Rex bioreactors
| Autor: | Christine Gagliardi, Aaron E. Foster, Mariam Khalil |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cancer Research T-Lymphocytes Immunology Cell Cell Culture Techniques Cell- and Tissue-Based Therapy Lymphocyte Activation Peripheral blood mononuclear cell Permeability Cell therapy 03 medical and health sciences Transduction (genetics) Bioreactors 0302 clinical medicine Immune system Transduction Genetic medicine Bioreactor Humans Immunology and Allergy Cells Cultured Genetics (clinical) Cell Proliferation Transplantation Receptors Chimeric Antigen Organisms Genetically Modified Tissue Engineering biology Chemistry Interleukin Cell Differentiation Equipment Design Cell Biology Cell biology 030104 developmental biology medicine.anatomical_structure Oncology 030220 oncology & carcinogenesis Leukocytes Mononuclear biology.protein Gases Antibody |
| Zdroj: | Cytotherapy. 21:1246-1257 |
| ISSN: | 1465-3249 |
| Popis: | Background Gas Permeable Rapid Expansion (G-Rex) bioreactors have been shown to efficiently expand immune cells intended for therapeutic use, but do not address the complexity of the viral transduction step required for many engineered T-cell products. Here we demonstrate a novel method for transduction of activated T cells with Vectofusin-1 reagent. Transduction is accomplished in suspension, in G-Rex bioreactors. The simplified transduction step is integrated into a streamlined process that uses a single bioreactor with limited operator intervention. Methods Peripheral blood mononuclear cells (PBMCs) from healthy donors were thawed, washed and activated with soluble anti-CD3 and anti-CD28 antibodies either in cell culture bags or in G-Rex bioreactors. Cells were cultured in TexMACS GMP medium with interleukin (IL)-7 and IL-15 and transduced with RetroNectin in bags or Vectorfusin-1 in the G-Rex. Total viable cell number, fold expansion, viability, transduction efficiency, phenotype and function were compared between the two processes. Results The simplified process uses a single vessel from activation through harvest and achieves 56% transduction with 29-fold expansion in 11 days. The cells generated in the simplified process do not differ from cells produced in the conventional bag-based process functionally or phenotypically. Discussion This study demonstrates that T cells can be transduced in suspension. Further, the conventional method of generating engineered T cells in bags for clinical use can be streamlined to a much simpler, less-expensive process without compromising the quality or function of the cell product. |
| Databáze: | OpenAIRE |
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