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
相关论文
共 37 条
  • [21] Interrater reliability: the kappa statistic
    McHugh, Mary L.
    [J]. BIOCHEMIA MEDICA, 2012, 22 (03) : 276 - 282
  • [22] Metabolic active-high density VERO cell cultures on microcarriers following apoptosis prevention by galactose/glutamine feeding
    Mendonça, RZ
    Arrózio, SJ
    Antoniazzi, MM
    Ferreira, JMC
    Pereira, CA
    [J]. JOURNAL OF BIOTECHNOLOGY, 2002, 97 (01) : 13 - 22
  • [23] PROPAGATION OF POLIOVIRUS IN MICROCARRIER CULTURES OF 3 MONKEY KIDNEY-CELL LINES
    MERED, B
    ALBRECHT, P
    HOPPS, HE
    PETRICCIANI, JC
    SALK, J
    [J]. JOURNAL OF BIOLOGICAL STANDARDIZATION, 1981, 9 (02): : 137 - 145
  • [24] NILSSON K, 1989, BIOTECHNOL GENET ENG, V6, P403
  • [25] Patterson M K Jr, 1979, Methods Enzymol, V58, P141
  • [26] APPLICATION OF QUANTITATIVE IMAGE-ANALYSIS TO A MAMMALIAN-CELL LINE GROWN ON MICROCARRIERS
    PONS, MN
    WAGNER, A
    VIVIER, H
    MARC, A
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 1992, 40 (01) : 187 - 193
  • [27] Automated multi-parameter measurement of cardiomyocytes dynamics with digital holographic microscopy
    Rappaz, Benjamin
    Moon, Inkyu
    Yi, Faliu
    Javidi, Bahram
    Marquet, Pierre
    Turcatti, Gerardo
    [J]. OPTICS EXPRESS, 2015, 23 (10): : 13333 - 13347
  • [28] Online monitoring of microcarrier based fibroblast cultivations with in situ microscopy
    Rudolph, Guido
    Lindner, Patrick
    Gierse, Alexander
    Bluma, Arne
    Martinez, Geovanni
    Hitzmann, Bernd
    Scheper, Thomas
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 2008, 99 (01) : 136 - 145
  • [29] Ear mesenchymal stem cells: An efficient adult multipotent cell population fit for rapid and scalable expansion
    Sart, Sebastien
    Schneider, Yves-Jacques
    Agathos, Spiros N.
    [J]. JOURNAL OF BIOTECHNOLOGY, 2009, 139 (04) : 291 - 299
  • [30] Stirred bioreactors for the expansion of adult pancreatic stem cells
    Serra, Margarida
    Brito, Catarina
    Leite, Sofia B.
    Gorjup, Erwin
    von Briesen, Hagen
    Carrondo, Manuel J. T.
    Alves, Paula M.
    [J]. ANNALS OF ANATOMY-ANATOMISCHER ANZEIGER, 2009, 191 (01) : 104 - 115