High throughput intracellular delivery by viscoelastic mechanoporation
Abstract Brief pulses of electric field (electroporation) and/or tensile stress (mechanoporation) have been used to reversibly permeabilize the plasma membrane of mammalian cells and deliver materials to the cytosol. However, electroporation can be harmful to cells, while efficient mechanoporation s...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-44447-w |
| _version_ | 1797363048457961472 |
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| author | Derin Sevenler Mehmet Toner |
| author_facet | Derin Sevenler Mehmet Toner |
| author_sort | Derin Sevenler |
| collection | DOAJ |
| description | Abstract Brief pulses of electric field (electroporation) and/or tensile stress (mechanoporation) have been used to reversibly permeabilize the plasma membrane of mammalian cells and deliver materials to the cytosol. However, electroporation can be harmful to cells, while efficient mechanoporation strategies have not been scalable due to the use of narrow constrictions or needles which are susceptible to clogging. Here we report a high throughput approach to mechanoporation in which the plasma membrane is stretched and reversibly permeabilized by viscoelastic fluid forces within a microfluidic chip without surface contact. Biomolecules are delivered directly to the cytosol within seconds at a throughput exceeding 250 million cells per minute. Viscoelastic mechanoporation is compatible with a variety of biomolecules including proteins, RNA, and CRISPR-Cas9 ribonucleoprotein complexes, as well as a range of cell types including HEK293T cells and primary T cells. Altogether, viscoelastic mechanoporation appears feasible for contact-free permeabilization and delivery of biomolecules to mammalian cells ex vivo. |
| first_indexed | 2024-03-08T16:16:15Z |
| format | Article |
| id | doaj.art-54a9208d72b145fe8d915ead8cf36e37 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-08T16:16:15Z |
| publishDate | 2024-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-54a9208d72b145fe8d915ead8cf36e372024-01-07T12:35:01ZengNature PortfolioNature Communications2041-17232024-01-0115111210.1038/s41467-023-44447-wHigh throughput intracellular delivery by viscoelastic mechanoporationDerin Sevenler0Mehmet Toner1Center for Engineering in Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical SchoolCenter for Engineering in Medicine and Surgery, Department of Surgery, Massachusetts General Hospital, Harvard Medical SchoolAbstract Brief pulses of electric field (electroporation) and/or tensile stress (mechanoporation) have been used to reversibly permeabilize the plasma membrane of mammalian cells and deliver materials to the cytosol. However, electroporation can be harmful to cells, while efficient mechanoporation strategies have not been scalable due to the use of narrow constrictions or needles which are susceptible to clogging. Here we report a high throughput approach to mechanoporation in which the plasma membrane is stretched and reversibly permeabilized by viscoelastic fluid forces within a microfluidic chip without surface contact. Biomolecules are delivered directly to the cytosol within seconds at a throughput exceeding 250 million cells per minute. Viscoelastic mechanoporation is compatible with a variety of biomolecules including proteins, RNA, and CRISPR-Cas9 ribonucleoprotein complexes, as well as a range of cell types including HEK293T cells and primary T cells. Altogether, viscoelastic mechanoporation appears feasible for contact-free permeabilization and delivery of biomolecules to mammalian cells ex vivo.https://doi.org/10.1038/s41467-023-44447-w |
| spellingShingle | Derin Sevenler Mehmet Toner High throughput intracellular delivery by viscoelastic mechanoporation Nature Communications |
| title | High throughput intracellular delivery by viscoelastic mechanoporation |
| title_full | High throughput intracellular delivery by viscoelastic mechanoporation |
| title_fullStr | High throughput intracellular delivery by viscoelastic mechanoporation |
| title_full_unstemmed | High throughput intracellular delivery by viscoelastic mechanoporation |
| title_short | High throughput intracellular delivery by viscoelastic mechanoporation |
| title_sort | high throughput intracellular delivery by viscoelastic mechanoporation |
| url | https://doi.org/10.1038/s41467-023-44447-w |
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