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. A medium size 20ml hollow fibre cartridge (C2011) offers 3000 cm2 of porous fibre surface for cell attachment and will support continuous post-confluent growth of 10e9 cells. Below you can find examples and publications.

  • Increase exosome yield by up to 100-fold compared to flask culture
  • Obtain very concentrated exosomes/EVs in small volumes of supernatant
  • High cell density bioreactor culture offers more physiological 3D growth 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 enabling periodic harvesting of concentrated EVs in 20ml syringe volumes. No passaging is required. Batch production in multiple flasks is avoided. Click on the article below to learn more about this hghly productive continuous culture system.

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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