A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells
Single-cell cloning (SCC) is a critical step in generating monoclonal cell lines, which are widely used as in vitro models and for producing proteins with high reproducibility for research and the production of therapeutic drugs. In monoclonal cell line generation, the development time can be shorte...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-06-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/6/1482 |
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| author | Chuan-Feng Yeh Ching-Hui Lin Hao-Chen Chang Chia-Yu Tang Pei-Tzu Lai Chia-Hsien Hsu |
| author_facet | Chuan-Feng Yeh Ching-Hui Lin Hao-Chen Chang Chia-Yu Tang Pei-Tzu Lai Chia-Hsien Hsu |
| author_sort | Chuan-Feng Yeh |
| collection | DOAJ |
| description | Single-cell cloning (SCC) is a critical step in generating monoclonal cell lines, which are widely used as in vitro models and for producing proteins with high reproducibility for research and the production of therapeutic drugs. In monoclonal cell line generation, the development time can be shortened by validating the monoclonality of the cloned cells. However, the validation process currently requires specialized equipment that is not readily available in general biology laboratories. Here, we report a disposable SCC device, in which single cells can be isolated, validated, and expanded to form monoclonal cell colonies using conventional micropipettes and microscopes. The monoclonal cells can be selectively transferred from the SCC chip to conventional culture plates, using a tissue puncher. Using the device, we demonstrated that monoclonal colonies of actin-GFP (green fluorescent protein) plasmid-transfected A549 cells could be formed in the device within nine days and subsequently transferred to wells in plates for further expansion. This approach offers a cost-effective alternative to the use of specialized equipment for monoclonal cell generation. |
| first_indexed | 2024-03-10T19:04:31Z |
| format | Article |
| id | doaj.art-c652895885684f2b8b46b70bd7bf598e |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T19:04:31Z |
| publishDate | 2020-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-c652895885684f2b8b46b70bd7bf598e2023-11-20T04:12:37ZengMDPI AGCells2073-44092020-06-0196148210.3390/cells9061482A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal CellsChuan-Feng Yeh0Ching-Hui Lin1Hao-Chen Chang2Chia-Yu Tang3Pei-Tzu Lai4Chia-Hsien Hsu5Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, TaiwanInstitute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, TaiwanInstitute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, TaiwanInstitute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, TaiwanInstitute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, TaiwanInstitute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, TaiwanSingle-cell cloning (SCC) is a critical step in generating monoclonal cell lines, which are widely used as in vitro models and for producing proteins with high reproducibility for research and the production of therapeutic drugs. In monoclonal cell line generation, the development time can be shortened by validating the monoclonality of the cloned cells. However, the validation process currently requires specialized equipment that is not readily available in general biology laboratories. Here, we report a disposable SCC device, in which single cells can be isolated, validated, and expanded to form monoclonal cell colonies using conventional micropipettes and microscopes. The monoclonal cells can be selectively transferred from the SCC chip to conventional culture plates, using a tissue puncher. Using the device, we demonstrated that monoclonal colonies of actin-GFP (green fluorescent protein) plasmid-transfected A549 cells could be formed in the device within nine days and subsequently transferred to wells in plates for further expansion. This approach offers a cost-effective alternative to the use of specialized equipment for monoclonal cell generation.https://www.mdpi.com/2073-4409/9/6/1482microfluidicssingle-cell cloningmonoclonal cell lines |
| spellingShingle | Chuan-Feng Yeh Ching-Hui Lin Hao-Chen Chang Chia-Yu Tang Pei-Tzu Lai Chia-Hsien Hsu A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells Cells microfluidics single-cell cloning monoclonal cell lines |
| title | A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells |
| title_full | A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells |
| title_fullStr | A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells |
| title_full_unstemmed | A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells |
| title_short | A Microfluidic Single-Cell Cloning (SCC) Device for the Generation of Monoclonal Cells |
| title_sort | microfluidic single cell cloning scc device for the generation of monoclonal cells |
| topic | microfluidics single-cell cloning monoclonal cell lines |
| url | https://www.mdpi.com/2073-4409/9/6/1482 |
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