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Cell confluency analysis on microcarriers by micro-flow imaging

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Abstract

The productivity of cell culture-derived vaccines grown in anchorage-dependent animal cells is limited by bioreactor surface area. One way to increase the available surface area is by growing cells as monolayers on small spheres called microcarriers, which are approximately 100–250 μm in diameter. In order for microcarrier-based cell culture to be a success, it is important to understand the kinetics of cell growth on the microcarriers. Micro-flow imaging (MFI) is a simple and powerful technique that captures images and analyzes samples as they are drawn through a precision flow cell. In addition to providing size distribution and defect frequency data to compare microcarrier lots, MFI was used to generate hundreds of images to determine cell coverage and confluency on microcarriers. Same-day manual classification of these images provided upstream cell culture teams with actionable data that informed in-process decision making (e.g. time of infection). Additionally, an automated cell coverage algorithm was developed to increase the speed and throughput of the analyses.

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Acknowledgments

The authors thank our colleagues in Vaccine Bioprocess Research and Development and Global Vaccines and Biologics Commercialization for samples and technical assistance.

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Authors and Affiliations

  1. Vaccine Analytical Development, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA, 19486, USA

    Christopher J. Farrell, Tim Culp & Van M. Hoang

  2. Eurofins Lancaster Laboratories Professional Scientific Services, Lancaster, PA, USA

    Stephanie M. Cicalese

  3. Vaccine Drug Product Development, Merck & Co., Inc., West Point, PA, USA

    Harrison B. Davis

  4. Applied Mathematics and Modeling, Merck & Co., Inc., Rahway, NJ, USA

    Belma Dogdas & Tosha Shah

Authors
  1. Christopher J. Farrell
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  2. Stephanie M. Cicalese
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  3. Harrison B. Davis
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  4. Belma Dogdas
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  5. Tosha Shah
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  6. Tim Culp
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  7. Van M. Hoang
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Correspondence to Christopher J. Farrell.

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Farrell, C.J., Cicalese, S.M., Davis, H.B. et al. Cell confluency analysis on microcarriers by micro-flow imaging. Cytotechnology 68, 2469–2478 (2016). https://doi.org/10.1007/s10616-016-9967-0

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  • DOI: https://doi.org/10.1007/s10616-016-9967-0

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