Dataset Information

Isolation and Characterization of small EVs from CHO-K1 cell cultures

ABSTRACT: Chinese hamster ovary (CHO) cells are widely used host cells for recombinant protein production and currently the most commonly utilized mammalian organism in large scale biopharmaceutical production. Since the discovery of exosomes as a new type of small extracellular vesicles (EVs) by Johnstone et al., interest in EV research greatly increased in recent years. However, they yet pose a blank space in CHO research. Exosomes are 30 – 150 nm small vesicles, that derive from the endosomal network and can therefore be distinguished from plasma membrane-shed microvesicles (100 – 1000 nm in diameter) and apoptotic vesicles (50 – 5000 nm), which are secluded over the course of programmed cell death. It turned out exosomes are not only vehicles of cellular waste disposal, as was initially assumed, but also a conserved mechanism of cellular communication. This work aims to outline possible separation techniques followed by a differential proteomic and transcriptomic characterization of CHO EVs over the course of a bioreactor batch cultivation. Therefore, a protocol yielding sEVs with a strong exosomal marker enrichment is compared with HCP, lEVs and whole cell lysate (WCL) from the same batch process. This may allow for further studies in this field to have reference data for evaluation of exosome isolation techniques, separation purity and CHO EV composition in general.

INSTRUMENT(S):

ORGANISM(S): Cricetulus Griseus (chinese Hamster) (cricetulus Barabensis Griseus)

TISSUE(S): Cell Suspension Culture, Permanent Cell Line Cell

SUBMITTER: Louise Schelletter