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Cytotechnology. 2016 Dec;68(6):2469-2478. Epub 2016 May 14.

Cell confluency analysis on microcarriers by micro-flow imaging.

Author information

1
Vaccine Analytical Development, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA, 19486, USA. christopher.farrell@merck.com.
2
Eurofins Lancaster Laboratories Professional Scientific Services, Lancaster, PA, USA.
3
Vaccine Drug Product Development, Merck & Co., Inc., West Point, PA, USA.
4
Applied Mathematics and Modeling, Merck & Co., Inc., Rahway, NJ, USA.
5
Vaccine Analytical Development, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA, 19486, USA.

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.

KEYWORDS:

Confluency; Mammalian cell culture; Micro-flow Imaging; Microcarriers

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