Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing
There is a paradigm shift in biomanufacturing toward continuous bioprocessing but cell-based manufacturing using adherent and suspension cultures, including microcarriers, hydrogel microparticles, and 3D cell aggregates, remains challenging due to the lack of efficient in-line bioprocess monitoring...
| Main Authors: | , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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2024
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| Online Access: | https://hdl.handle.net/10356/174671 |
| _version_ | 1826115484673638400 |
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| author | Gong, Lingyan He, Linwei Lu, Nan Petchakup, Chayakorn Li, Holden King Ho Tay, Chor Yong Hou, Han Wei |
| author2 | School of Mechanical and Aerospace Engineering |
| author_facet | School of Mechanical and Aerospace Engineering Gong, Lingyan He, Linwei Lu, Nan Petchakup, Chayakorn Li, Holden King Ho Tay, Chor Yong Hou, Han Wei |
| author_sort | Gong, Lingyan |
| collection | NTU |
| description | There is a paradigm shift in biomanufacturing toward continuous bioprocessing but cell-based manufacturing using adherent and suspension cultures, including microcarriers, hydrogel microparticles, and 3D cell aggregates, remains challenging due to the lack of efficient in-line bioprocess monitoring and cell harvesting tools. Herein, a novel label-free microfluidic platform for high throughput (≈50 particles/sec) impedance bioanalysis of biomass, cell viability, and stem cell differentiation at single particle resolution is reported. The device is integrated with a real-time piezo-actuated particle sorter based on user-defined multi-frequency impedance signatures. Biomass profiling of Cytodex-3 microcarriers seeded with adipose-derived mesenchymal stem cells (ADSCs) is first performed to sort well-seeded or confluent microcarriers for downstream culture or harvesting, respectively. Next, impedance-based isolation of microcarriers with osteogenic differentiated ADSCs is demonstrated, which is validated with a twofold increase of calcium content in sorted ADSCs. Impedance profiling of heterogenous ADSCs-encapsulated hydrogel (alginate) microparticles and 3D ADSC aggregate mixtures is also performed to sort particles with high biomass and cell viability to improve cell quality. Overall, the scalable microfluidic platform technology enables in-line sample processing from bioreactors directly and automated analysis of cell quality attributes to maximize cell yield and improve the control of cell quality in continuous cell-based manufacturing. |
| first_indexed | 2024-10-01T03:55:54Z |
| format | Journal Article |
| id | ntu-10356/174671 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:55:54Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | ntu-10356/1746712024-04-13T16:49:12Z Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing Gong, Lingyan He, Linwei Lu, Nan Petchakup, Chayakorn Li, Holden King Ho Tay, Chor Yong Hou, Han Wei School of Mechanical and Aerospace Engineering School of Materials Science and Engineering Lee Kong Chian School of Medicine (LKCMedicine) Nanyang Environment and Water Research Institute Environmental Chemistry and Materials Centre Engineering Cell-based manufacturing Hydrogel microparticles There is a paradigm shift in biomanufacturing toward continuous bioprocessing but cell-based manufacturing using adherent and suspension cultures, including microcarriers, hydrogel microparticles, and 3D cell aggregates, remains challenging due to the lack of efficient in-line bioprocess monitoring and cell harvesting tools. Herein, a novel label-free microfluidic platform for high throughput (≈50 particles/sec) impedance bioanalysis of biomass, cell viability, and stem cell differentiation at single particle resolution is reported. The device is integrated with a real-time piezo-actuated particle sorter based on user-defined multi-frequency impedance signatures. Biomass profiling of Cytodex-3 microcarriers seeded with adipose-derived mesenchymal stem cells (ADSCs) is first performed to sort well-seeded or confluent microcarriers for downstream culture or harvesting, respectively. Next, impedance-based isolation of microcarriers with osteogenic differentiated ADSCs is demonstrated, which is validated with a twofold increase of calcium content in sorted ADSCs. Impedance profiling of heterogenous ADSCs-encapsulated hydrogel (alginate) microparticles and 3D ADSC aggregate mixtures is also performed to sort particles with high biomass and cell viability to improve cell quality. Overall, the scalable microfluidic platform technology enables in-line sample processing from bioreactors directly and automated analysis of cell quality attributes to maximize cell yield and improve the control of cell quality in continuous cell-based manufacturing. Ministry of Education (MOE) Submitted/Accepted version This research was supported by Singapore Ministry of Education (MOE) AcRF Tier 2 (MOE-T2EP30120-0004) and MOE AcRF Tier 1 (RG29/23) awarded to H.W.H. L.G. acknowledges support for the NTU PhD Research Scholarship. 2024-04-07T07:26:53Z 2024-04-07T07:26:53Z 2024 Journal Article Gong, L., He, L., Lu, N., Petchakup, C., Li, H. K. H., Tay, C. Y. & Hou, H. W. (2024). Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing. Advanced Healthcare Materials. https://dx.doi.org/10.1002/adhm.202304529 2192-2640 https://hdl.handle.net/10356/174671 10.1002/adhm.202304529 38465888 2-s2.0-85187943696 en MOE-T2EP30120-0004 RG29/23 Advanced Healthcare Materials © 2024 Wiley-VCH GmbH. All rights reserved. This article may be downloaded for personal use only. Any other use requires prior permission of the copyright holder. The Version of Record is available online at http://doi.org/10.1002/adhm.202304529. application/pdf application/pdf |
| spellingShingle | Engineering Cell-based manufacturing Hydrogel microparticles Gong, Lingyan He, Linwei Lu, Nan Petchakup, Chayakorn Li, Holden King Ho Tay, Chor Yong Hou, Han Wei Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing |
| title | Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing |
| title_full | Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing |
| title_fullStr | Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing |
| title_full_unstemmed | Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing |
| title_short | Label-free single microparticles and cell aggregates sorting in continuous cell-based manufacturing |
| title_sort | label free single microparticles and cell aggregates sorting in continuous cell based manufacturing |
| topic | Engineering Cell-based manufacturing Hydrogel microparticles |
| url | https://hdl.handle.net/10356/174671 |
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