Easy Scale-Up of Exosome/EVs Production

  • Increase exosome yield by up to 100-fold compared to flask culture
  • Obtain concentrated exosomes/EVs
  • Serum-free high cell density bioreactor conditions

Standardizing production of large quantities of exosomes/ EV’s

A FiberCell hollow fibre bioreactor enables the standardized production of large amounts of ready-concentrated 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, EVs accumulate in the extra-capillary space and can be periodically harvested over a continuous culture period of weeks or even months.

Cells are not subjected to shear stress and are resistant to apoptosis in the perfused, capillarized cartridge resulting in clean EV harvests that are free of apoptotic bodies. Also the 20 Kd fibre cut-off blocks contamination from any bovine EVs due to serum in recirculating media.

To our knowledge, in hollow fibre culture, stem cells do not differentiate, resulting in linear production over time with the advantage of phenotypic stability.

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:

Scalable, cGMP-compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL-15/lactadherin complexes: Watson,D.C. et al.; J Extracell Vesicles. 2018 Feb 28;7(1) [open acccess]

Use of a Hollow Fiber Bioreactor to Collect Extracellular Vesicles from Cells in Culture: Yan IK et al.; Methods Mol Biol. 2018;1740:35-41 [abstract]

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
CDM-HD Serum substitute for high cell density bioreactor (12 week run) 4

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