Quantification of the dynamics of population heterogeneities in CHO cultures with stably integrated fluorescent markers.

Analytical and Bioanalytical Chemistry
Johannes MöllerUwe Jandt

Abstract

Cell population heterogeneities and their changes in mammalian cell culture processes are still not well characterized. In this study, the formation and dynamics of cell population heterogeneities were investigated with flow cytometry and stably integrated fluorescent markers based on the lentiviral gene ontology (LeGO) vector system. To achieve this, antibody-producing CHO cells were transduced with different LeGO vectors to stably express single or multiple fluorescent proteins. This enables the tracking of the transduced populations and is discussed in two case studies from the field of bioprocess engineering: In case study I, cells were co-transduced to express red, green, and blue fluorescent proteins and the development of sub-populations and expression heterogeneities were investigated in high passage cultivations (total 130 days). The formation of a fast-growing and more productive population was observed with a simultaneous increase in cell density and product titer. In case study II, different preculture growth phases and their influence on the population dynamics were investigated in mixed batch cultures with flow cytometry (offline and automated). Four cell line derivatives, each expressing a different fluorescent p...Continue Reading

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Citations

Jan 21, 2021·Bioprocess and Biosystems Engineering·André MoserJohannes Möller
Mar 16, 2021·Engineering in Life Sciences·Johannes MöllerAn-Ping Zeng

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Methods Mentioned

BETA
flow
fluorescence microscopy
flow cytometry
biosensors
glycosylation
by flow cytometry
fluorescence-activated cell sorting

Software Mentioned

color View
Addgene
MATLAB
CellˆP
GIMP
LeGO

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