Easy Scale-Up of Exosome/EVs Production

  • Increase exosome yield by around 100-fold compared to flask culture
  • Obtain ready-concentrated exosomes/EVs
  • No need for serum starvation prior to collection

Standardizing production of large quantities of exosomes/ EV’s

A FiberCell hollow fibre bioreactor enables the standardized production of large amounts of ready-concentrated exosomes/EVs from cells cultured or co-cultured at high density: tumour and engineered cell lines, stem cells etc.. Retained by the 20 kd MWCO fibres, exosomes accumulate in the extra-capillary space. They can be easily harvested every few days over a period of weeks or even months.

Cells are resistant to apoptosis in the perfused, capillarized cartridge resulting in clean exosome harvests that are free of apoptotic bodies. Also the fibre cut-off very effectively stops contamination by any bovine exosomes present in the recirculating media.

To our knowledge, in hollow fibre culture, stem cells do not differentiate, remaining phenotypically stable.

FiberCell® HFBR easily outperforms T-225 flasks

Flasks were split 1:5 in DMEM/10% FBS until a total of 130 T225 flasks were obtained. DMEM/10% FBS was replaced with DMEM alone and exosomes collected after 2 days.

5X10^8 adipose-derived mesenchymal stem cells were seeded into the hollow fibre bioreactor (C2011 cartridge) and harvests initiated after one week of culture. Medium used in the hollow fibre bioreactors was DMEM/10% FBS in the circulating medium, DMEM without serum in the extra-capillary space.

Total exosomes collected from two HFBR under these conditions equalled that from 3600x T225 flasks in a total volume of 360 mL.

Collection Volume (ml) Total Exosome Protein (mg) Total Exosome Particles (1010)
HFBR
  Cartridge #1 240 11.82 95.8
  Cartridge #2 120 14.45 326.9
 T-225
 130 flasks 4000 0.9 1.6

Cartridge #1 – 7 weeks, weekly collection
Cartridge #2 – 4 weeks, 6 collections

Total medium consumed:
hollow fibre: 7 L per run
flasks: 24 L

References:

Clinical Scale Production and Wound Healing Activity of Human Adipose Derived Mesenchymal Stem Cell Extracellular Vesicles from a Hollow Fiber Bioreactor. [Fibercell Poster ISEV2017]

Efficient production and enhanced tumor delivery of engineered extracellular vesicles: Watson DC et al.; Biomaterials 105, 2016: 195-205 [Open Access]

Starter kit for exosome production

Cat.# Description Qty
P3202 FiberCell® Systems Duet 1
C2011 High Flux PS Cartridge 20 kd (3000 cm2 – 20 ml) 1
A – – – 33mm, 38mm or 45mm Reservoir Cap 1

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