Optimized conditions for gene transduction into primary immune cells using viral vectors
Abstract Chimeric antigen receptor (CAR) T cell therapy has emerged as a promising modality for anti-cancer treatment. Its efficacy is quite remarkable in hematological tumors. Owing to their excellent clinical results, gene- modified cell therapies, including T cells, natural killer (NK) cells, and...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-39597-2 |
| _version_ | 1827875528787361792 |
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| author | Yeongrin Kim Da Yeon Lee Ji U Choi Jin Song Park So Myoung Lee Chung Hyo Kang Chi Hoon Park |
| author_facet | Yeongrin Kim Da Yeon Lee Ji U Choi Jin Song Park So Myoung Lee Chung Hyo Kang Chi Hoon Park |
| author_sort | Yeongrin Kim |
| collection | DOAJ |
| description | Abstract Chimeric antigen receptor (CAR) T cell therapy has emerged as a promising modality for anti-cancer treatment. Its efficacy is quite remarkable in hematological tumors. Owing to their excellent clinical results, gene- modified cell therapies, including T cells, natural killer (NK) cells, and macrophages, are being actively studied in both academia and industry. However, the protocol to make CAR immune cells is too complicated, so it is still unclear how to efficiently produce the potent CAR immune cells. To manufacture effective CAR immune cells, we need to be aware of not only how to obtain highly infective viral particles, but also how to transduce CAR genes into immune cells. In this paper, we provide detailed information on spinoculation, which is one of the best known protocols to transduce genes into immune cells, in a methodological view. Our data indicate that gene transduction is significantly dependent on speed and duration of centrifugation, concentration and number of viral particles, the concentration of polybrene, and number of infected immune cells. In addition, we investigated on the optimal polyethylene glycol (PEG) solution to concentrate the viral supernatant and the optimized DNA ratios transfected into 293T cells to produce high titer of viral particles. This study provides useful information for practical production of the gene-modified immune cells using viral vectors. |
| first_indexed | 2024-03-12T17:09:13Z |
| format | Article |
| id | doaj.art-60f107242f88406f9df7e3ef154b596f |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-12T17:09:13Z |
| publishDate | 2023-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-60f107242f88406f9df7e3ef154b596f2023-08-06T11:12:15ZengNature PortfolioScientific Reports2045-23222023-07-0113111210.1038/s41598-023-39597-2Optimized conditions for gene transduction into primary immune cells using viral vectorsYeongrin Kim0Da Yeon Lee1Ji U Choi2Jin Song Park3So Myoung Lee4Chung Hyo Kang5Chi Hoon Park6Bio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyBio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyBio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyBio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyBio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyBio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyBio and Drug Discovery Division, Korea Research Institute of Chemical TechnologyAbstract Chimeric antigen receptor (CAR) T cell therapy has emerged as a promising modality for anti-cancer treatment. Its efficacy is quite remarkable in hematological tumors. Owing to their excellent clinical results, gene- modified cell therapies, including T cells, natural killer (NK) cells, and macrophages, are being actively studied in both academia and industry. However, the protocol to make CAR immune cells is too complicated, so it is still unclear how to efficiently produce the potent CAR immune cells. To manufacture effective CAR immune cells, we need to be aware of not only how to obtain highly infective viral particles, but also how to transduce CAR genes into immune cells. In this paper, we provide detailed information on spinoculation, which is one of the best known protocols to transduce genes into immune cells, in a methodological view. Our data indicate that gene transduction is significantly dependent on speed and duration of centrifugation, concentration and number of viral particles, the concentration of polybrene, and number of infected immune cells. In addition, we investigated on the optimal polyethylene glycol (PEG) solution to concentrate the viral supernatant and the optimized DNA ratios transfected into 293T cells to produce high titer of viral particles. This study provides useful information for practical production of the gene-modified immune cells using viral vectors.https://doi.org/10.1038/s41598-023-39597-2 |
| spellingShingle | Yeongrin Kim Da Yeon Lee Ji U Choi Jin Song Park So Myoung Lee Chung Hyo Kang Chi Hoon Park Optimized conditions for gene transduction into primary immune cells using viral vectors Scientific Reports |
| title | Optimized conditions for gene transduction into primary immune cells using viral vectors |
| title_full | Optimized conditions for gene transduction into primary immune cells using viral vectors |
| title_fullStr | Optimized conditions for gene transduction into primary immune cells using viral vectors |
| title_full_unstemmed | Optimized conditions for gene transduction into primary immune cells using viral vectors |
| title_short | Optimized conditions for gene transduction into primary immune cells using viral vectors |
| title_sort | optimized conditions for gene transduction into primary immune cells using viral vectors |
| url | https://doi.org/10.1038/s41598-023-39597-2 |
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