Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells
Summary: Human embryonic kidney cells HEK293 can be used for the production of therapeutic glycoproteins requiring human post-translational modifications. High cell density perfusion processes are advantageous for such production but are challenging due to the shear sensitivity of HEK293 cells. To u...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-11-01
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| Series: | iScience |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004220308452 |
| _version_ | 1811222207888621568 |
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| author | Caijuan Zhan Gholamreza Bidkhori Hubert Schwarz Magdalena Malm Aman Mebrahtu Ray Field Christopher Sellick Diane Hatton Paul Varley Adil Mardinoglu Johan Rockberg Veronique Chotteau |
| author_facet | Caijuan Zhan Gholamreza Bidkhori Hubert Schwarz Magdalena Malm Aman Mebrahtu Ray Field Christopher Sellick Diane Hatton Paul Varley Adil Mardinoglu Johan Rockberg Veronique Chotteau |
| author_sort | Caijuan Zhan |
| collection | DOAJ |
| description | Summary: Human embryonic kidney cells HEK293 can be used for the production of therapeutic glycoproteins requiring human post-translational modifications. High cell density perfusion processes are advantageous for such production but are challenging due to the shear sensitivity of HEK293 cells. To understand the impact of hollow filter cell separation devices, cells were cultured in bioreactors operated with tangential flow filtration (TFF) or alternating tangential flow filtration (ATF) at various flow rates. The average theoretical velocity profile in these devices showed a lower shear stress for ATF by a factor 0.637 compared to TFF. This was experimentally validated and, furthermore, transcriptomic evaluation provided insights into the underlying cellular processes. High shear caused cellular stress leading to apoptosis by three pathways, i.e. endoplasmic reticulum stress, cytoskeleton reorganization, and extrinsic signaling pathways. Positive effects of mild shear stress were observed, with increased recombinant erythropoietin production and increased gene expression associated with transcription and protein phosphorylation. |
| first_indexed | 2024-04-12T08:12:04Z |
| format | Article |
| id | doaj.art-3527cbee461a49d0a4b65321f003c3e5 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-04-12T08:12:04Z |
| publishDate | 2020-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-3527cbee461a49d0a4b65321f003c3e52022-12-22T03:40:56ZengElsevieriScience2589-00422020-11-012311101653Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human CellsCaijuan Zhan0Gholamreza Bidkhori1Hubert Schwarz2Magdalena Malm3Aman Mebrahtu4Ray Field5Christopher Sellick6Diane Hatton7Paul Varley8Adil Mardinoglu9Johan Rockberg10Veronique Chotteau11KTH - Cell Technology Group (CETEG), Department of Industrial Biotechnology, 106 91, Stockholm, Sweden; Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, Sweden; AdBIOPRO, Competence Centre for Advanced Bioproduction by Continuous Processing, Stockholm, SwedenScience for Life Laboratory, KTH - Royal Institute of Technology, 171 21, Stockholm, SwedenKTH - Cell Technology Group (CETEG), Department of Industrial Biotechnology, 106 91, Stockholm, Sweden; Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, Sweden; AdBIOPRO, Competence Centre for Advanced Bioproduction by Continuous Processing, Stockholm, SwedenKTH - Royal Institute of Technology, Department of Protein Science, 106 91 Stockholm, Sweden; Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, SwedenKTH - Royal Institute of Technology, Department of Protein Science, 106 91 Stockholm, Sweden; Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, SwedenBioPharmaceutical Development, AstraZeneca, Cambridge, UKBioPharmaceutical Development, AstraZeneca, Cambridge, UKBioPharmaceutical Development, AstraZeneca, Cambridge, UKBioPharmaceutical Development, AstraZeneca, Cambridge, UKScience for Life Laboratory, KTH - Royal Institute of Technology, 171 21, Stockholm, SwedenKTH - Royal Institute of Technology, Department of Protein Science, 106 91 Stockholm, Sweden; Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, Sweden; AdBIOPRO, Competence Centre for Advanced Bioproduction by Continuous Processing, Stockholm, SwedenKTH - Cell Technology Group (CETEG), Department of Industrial Biotechnology, 106 91, Stockholm, Sweden; Wallenberg Centre for Protein Research (WCPR), 106 91 Stockholm, Sweden; AdBIOPRO, Competence Centre for Advanced Bioproduction by Continuous Processing, Stockholm, Sweden; Corresponding authorSummary: Human embryonic kidney cells HEK293 can be used for the production of therapeutic glycoproteins requiring human post-translational modifications. High cell density perfusion processes are advantageous for such production but are challenging due to the shear sensitivity of HEK293 cells. To understand the impact of hollow filter cell separation devices, cells were cultured in bioreactors operated with tangential flow filtration (TFF) or alternating tangential flow filtration (ATF) at various flow rates. The average theoretical velocity profile in these devices showed a lower shear stress for ATF by a factor 0.637 compared to TFF. This was experimentally validated and, furthermore, transcriptomic evaluation provided insights into the underlying cellular processes. High shear caused cellular stress leading to apoptosis by three pathways, i.e. endoplasmic reticulum stress, cytoskeleton reorganization, and extrinsic signaling pathways. Positive effects of mild shear stress were observed, with increased recombinant erythropoietin production and increased gene expression associated with transcription and protein phosphorylation.http://www.sciencedirect.com/science/article/pii/S2589004220308452BioengineeringBiophysicsCell Biology |
| spellingShingle | Caijuan Zhan Gholamreza Bidkhori Hubert Schwarz Magdalena Malm Aman Mebrahtu Ray Field Christopher Sellick Diane Hatton Paul Varley Adil Mardinoglu Johan Rockberg Veronique Chotteau Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells iScience Bioengineering Biophysics Cell Biology |
| title | Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells |
| title_full | Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells |
| title_fullStr | Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells |
| title_full_unstemmed | Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells |
| title_short | Low Shear Stress Increases Recombinant Protein Production and High Shear Stress Increases Apoptosis in Human Cells |
| title_sort | low shear stress increases recombinant protein production and high shear stress increases apoptosis in human cells |
| topic | Bioengineering Biophysics Cell Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004220308452 |
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