Cell confluency analysis on microcarriers by micro-flow imaging

被引:13
作者
Farrell, Christopher J. [1 ]
Cicalese, Stephanie M. [2 ]
Davis, Harrison B. [3 ]
Dogdas, Belma [4 ]
Shah, Tosha [4 ]
Culp, Tim [1 ]
Hoang, Van M. [1 ]
机构
[1] Merck & Co Inc, Vaccine Analyt Dev, 770 Sumneytown Pike, West Point, PA 19486 USA
[2] Eurofins Lancaster Labs, Profess Sci Serv, Lancaster, PA USA
[3] Merck & Co Inc, Vaccine Drug Prod Dev, West Point, PA USA
[4] Merck & Co Inc, Appl Math & Modeling, Rahway, NJ 07065 USA
关键词
Microcarriers; Micro-flow Imaging; Confluency; Mammalian cell culture; ANCHORAGE-DEPENDENT CELLS; EMBRYONIC STEM-CELLS; SUSPENSION-CULTURE; VIRUS; EXPANSION; PARTICLES; PRODUCTS; DISEASE; SYSTEM; GROWTH;
D O I
10.1007/s10616-016-9967-0
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 mu m 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.
引用
收藏
页码:2469 / 2478
页数:10
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