PUBLICATION Mammalian Cell Designers’ Laboratory

MISSION

Our mission is to engineer next-generation mammalian cell factories
for the production of complex biotherapeutics.
By integrating synthetic biology, genome engineering, and autonomous experimentation,
we aim to transform cell line development from empirical optimization into a predictive engineering discipline.

Journal Articles

Monitoring of autophagy in Chinese hamster ovary cells using flow cytometry
Author
Jae Seong Lee*, Gyun Min Lee‡
Journal
Methods
Vol
56(3)
Page
375-382
Year
2017~

Recently, autophagy, which is a degradative process, has drawn attention as an anti-cell death engineering target in addition to apoptosis in recombinant Chinese hamster ovary (rCHO) cell cultures for enhanced production of therapeutic proteins. Appropriate autophagy monitoring methods, that are suitable for long term CHO cell cultures, are necessary in order to investigate the culture conditions that affect the autophagy pathway and to select appropriate engineering targets for autophagy control. Herein, detailed protocols for autophagy monitoring methods based on flow cytometry are provided using the GFP-LC3-overexpressing CHO DG44 host cell line or MDC-like molecules in rCHO cells grown as an adherent culture with serum-containing medium or suspension culture with serum-free medium. Furthermore, combined with the apoptosis detection based on the Annexin V-PS interaction, the simultaneous detection of autophagy and apoptosis is also described. It is anticipated that the protocols described herein will assist in the fast, high throughput monitoring of autophagy that can support other existing autophagy assays.