Exosomes/EVs Production Scale-Up

Continuous production process for Exosomes / Microvesicles

Large amounts of EVs can be obtained from practically any type of cell, including mesenchymal stems cells, maintained over a few weeks at high density in a FiberCell Hollow Fibre Bioreactor. Below you can find examples and publications.

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

NTA Analysis of EV produced by Bone-marrow MSC in FiberCell Duet Bioreactor – Small Cartridge C2025D

mesenchymal stem cells production of exosomes in time-scalable process

Simple routine exosome collection process

A FiberCell Hollow Fibre Bioreactor provides a continuous production process requiring periodic harvesting of concentrated EVs in a syringe volume. No passaging is required. Batch production in multiple flasks is avoided.

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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