Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications
Microfluidic 3D tissue culture systems are attractive for in vitro drug testing applications due to the ability of these platforms to generate 3D tissue models and perform drug testing at a very small scale. However, the minute cell number and liquid volume impose significant technical challenges to...
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MDPI AG
2020-07-01
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| Online Access: | https://www.mdpi.com/2072-666X/11/7/669 |
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| author | Louis Jun Ye Ong Liang Zhu Gabriel Jenn Sern Tan Yi-Chin Toh |
| author_facet | Louis Jun Ye Ong Liang Zhu Gabriel Jenn Sern Tan Yi-Chin Toh |
| author_sort | Louis Jun Ye Ong |
| collection | DOAJ |
| description | Microfluidic 3D tissue culture systems are attractive for in vitro drug testing applications due to the ability of these platforms to generate 3D tissue models and perform drug testing at a very small scale. However, the minute cell number and liquid volume impose significant technical challenges to perform quantitative cell viability measurements using conventional colorimetric or fluorometric assays, such as MTS or Alamar Blue. Similarly, live-dead staining approaches often utilize metabolic dyes that typically label the cytoplasm of live cells, which makes it difficult to segment and count individual cells in compact 3D tissue cultures. In this paper, we present a quantitative image-based cell viability (QuantICV) assay technique that circumvents current challenges of performing the quantitative cell viability assay in microfluidic 3D tissue cultures. A pair of cell-impermeant nuclear dyes (EthD-1 and DAPI) were used to sequentially label the nuclei of necrotic and total cell populations, respectively. Confocal microscopy and image processing algorithms were employed to visualize and quantify the cell nuclei in the 3D tissue volume. The QuantICV assay was validated and showed good concordance with the conventional bulk MTS assay in static 2D and 3D tumor cell cultures. Finally, the QuantICV assay was employed as an on-chip readout to determine the differential dose responses of parental and metastatic 3D oral squamous cell carcinoma (OSCC) to Gefitinib in a microfluidic 3D culture device. This proposed technique can be useful in microfluidic cell cultures as well as in a situation where conventional cell viability assays are not available. |
| first_indexed | 2024-03-10T18:35:41Z |
| format | Article |
| id | doaj.art-7f918c382e6a4e72aa35f5cecef68f5f |
| institution | Directory Open Access Journal |
| issn | 2072-666X |
| language | English |
| last_indexed | 2024-03-10T18:35:41Z |
| publishDate | 2020-07-01 |
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| series | Micromachines |
| spelling | doaj.art-7f918c382e6a4e72aa35f5cecef68f5f2023-11-20T06:18:17ZengMDPI AGMicromachines2072-666X2020-07-0111766910.3390/mi11070669Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture ApplicationsLouis Jun Ye Ong0Liang Zhu1Gabriel Jenn Sern Tan2Yi-Chin Toh3Department of Biomedical Engineering, National University of Singapore, 4, Engineering Drive 3, E4-04-10, Singapore 117583, SingaporeDepartment of Biomedical Engineering, National University of Singapore, 4, Engineering Drive 3, E4-04-10, Singapore 117583, SingaporeDepartment of Biomedical Engineering, National University of Singapore, 4, Engineering Drive 3, E4-04-10, Singapore 117583, SingaporeDepartment of Biomedical Engineering, National University of Singapore, 4, Engineering Drive 3, E4-04-10, Singapore 117583, SingaporeMicrofluidic 3D tissue culture systems are attractive for in vitro drug testing applications due to the ability of these platforms to generate 3D tissue models and perform drug testing at a very small scale. However, the minute cell number and liquid volume impose significant technical challenges to perform quantitative cell viability measurements using conventional colorimetric or fluorometric assays, such as MTS or Alamar Blue. Similarly, live-dead staining approaches often utilize metabolic dyes that typically label the cytoplasm of live cells, which makes it difficult to segment and count individual cells in compact 3D tissue cultures. In this paper, we present a quantitative image-based cell viability (QuantICV) assay technique that circumvents current challenges of performing the quantitative cell viability assay in microfluidic 3D tissue cultures. A pair of cell-impermeant nuclear dyes (EthD-1 and DAPI) were used to sequentially label the nuclei of necrotic and total cell populations, respectively. Confocal microscopy and image processing algorithms were employed to visualize and quantify the cell nuclei in the 3D tissue volume. The QuantICV assay was validated and showed good concordance with the conventional bulk MTS assay in static 2D and 3D tumor cell cultures. Finally, the QuantICV assay was employed as an on-chip readout to determine the differential dose responses of parental and metastatic 3D oral squamous cell carcinoma (OSCC) to Gefitinib in a microfluidic 3D culture device. This proposed technique can be useful in microfluidic cell cultures as well as in a situation where conventional cell viability assays are not available.https://www.mdpi.com/2072-666X/11/7/6693D cell culturemicrofluidiccell viabilityquantitativetissue culture |
| spellingShingle | Louis Jun Ye Ong Liang Zhu Gabriel Jenn Sern Tan Yi-Chin Toh Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications Micromachines 3D cell culture microfluidic cell viability quantitative tissue culture |
| title | Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications |
| title_full | Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications |
| title_fullStr | Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications |
| title_full_unstemmed | Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications |
| title_short | Quantitative Image-Based Cell Viability (QuantICV) Assay for Microfluidic 3D Tissue Culture Applications |
| title_sort | quantitative image based cell viability quanticv assay for microfluidic 3d tissue culture applications |
| topic | 3D cell culture microfluidic cell viability quantitative tissue culture |
| url | https://www.mdpi.com/2072-666X/11/7/669 |
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