Volumetric imaging of human mesenchymal stem cells (hMSCs) for non-destructive quantification of 3D cell culture growth
The adoption of cell-based therapies into the clinic will require tremendous large-scale expansion to satisfy future demand, and bioreactor-microcarrier cultures are best suited to meet this challenge. The use of spherical microcarriers, however, precludes in-process visualization and monitoring of...
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| Veröffentlicht in: | PloS one 2023-03, Vol.18 (3), p.e0282298-e0282298 |
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| creator | Benavides, Oscar R Gibbs, Holly C White, Berkley P Kaunas, Roland Gregory, Carl A Walsh, Alex J Maitland, Kristen C |
| description | The adoption of cell-based therapies into the clinic will require tremendous large-scale expansion to satisfy future demand, and bioreactor-microcarrier cultures are best suited to meet this challenge. The use of spherical microcarriers, however, precludes in-process visualization and monitoring of cell number, morphology, and culture health. The development of novel expansion methods also motivates the advancement of analytical methods used to characterize these microcarrier cultures. A robust optical imaging and image-analysis assay to non-destructively quantify cell number and cell volume was developed. This method preserves 3D cell morphology and does not require membrane lysing, cellular detachment, or exogenous labeling. Complex cellular networks formed in microcarrier aggregates were imaged and analyzed in toto. Direct cell enumeration of large aggregates was performed in toto for the first time. This assay was successfully applied to monitor cellular growth of mesenchymal stem cells attached to spherical hydrogel microcarriers over time. Elastic scattering and fluorescence lightsheet microscopy were used to quantify cell volume and cell number at varying spatial scales. The presented study motivates the development of on-line optical imaging and image analysis systems for robust, automated, and non-destructive monitoring of bioreactor-microcarrier cell cultures. |
| doi_str_mv | 10.1371/journal.pone.0282298 |
| format | Article |
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The presented study motivates the development of on-line optical imaging and image analysis systems for robust, automated, and non-destructive monitoring of bioreactor-microcarrier cell cultures.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0282298</identifier><identifier>PMID: 36976801</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Aggregates ; Analysis ; Analytical methods ; Automation ; Bioreactors ; Cell culture ; Cell Culture Techniques - methods ; Cell Culture Techniques, Three Dimensional ; Cell morphology ; Cell number ; Cell Proliferation ; Cell size ; Cellular communication ; Cytology ; Elastic scattering ; Enumeration ; Evaluation ; Fluorescence ; Fluorescence microscopy ; Humans ; Hydrogels ; Image analysis ; Image processing ; Labeling ; Manufacturing ; Mesenchymal Stem Cells ; Methods ; Microscopy ; Morphology ; Pharmaceuticals ; Physical Sciences ; Product quality ; Real time ; Research and Analysis Methods ; Robustness ; Spectrum analysis ; Stem cells ; Visualization</subject><ispartof>PloS one, 2023-03, Vol.18 (3), p.e0282298-e0282298</ispartof><rights>Copyright: © 2023 Benavides et al. 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| source | Open Access: PubMed Central; PLoS; MEDLINE; Free E-Journal (出版社公開部分のみ）; DOAJ Directory of Open Access Journals; Free Full-Text Journals in Chemistry |
| subjects | Aggregates Analysis Analytical methods Automation Bioreactors Cell culture Cell Culture Techniques - methods Cell Culture Techniques, Three Dimensional Cell morphology Cell number Cell Proliferation Cell size Cellular communication Cytology Elastic scattering Enumeration Evaluation Fluorescence Fluorescence microscopy Humans Hydrogels Image analysis Image processing Labeling Manufacturing Mesenchymal Stem Cells Methods Microscopy Morphology Pharmaceuticals Physical Sciences Product quality Real time Research and Analysis Methods Robustness Spectrum analysis Stem cells Visualization |
| title | Volumetric imaging of human mesenchymal stem cells (hMSCs) for non-destructive quantification of 3D cell culture growth |
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