Production scale-up with different size cartridge modules

Continuous production process for Exomeres, Exosomes, Extracellular Vesicles, Microvesicles…

Some EV producers find themselves burdened with very large volumes of static flask supernatant to process which can lead to stressful bottlenecks in lab workflow. A FiberCell Hollow Fibre Bioreactor produces very concentrated EVs in a space- and time-efficient manner. Practically any type of cell, including mesenchymal stromal cells, can be cultured continuously at high density in this system. Yield increases of at least 40X compared to flask culture have been reported while the total amount of EVs produced depends on how many weeks the system is operated.

In addition, the 3D capillarised culture environment inside a hollow fibre bioreactor has been shown to be more representative of in vivo conditions compared to flask culture […more].

A Medium Size 20ml FiberCell cartridge (C2011) will support continuous post-confluent growth of 1E+9 cells. Further scale-up is straightforward without the need for new hardware.

  • 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

Small, Medium and Large Cartridge Modules available

Catering for pilot scale through to production, FiberCell cartridge modules are available in volumes up to 70ml with a fibre surface area of 1.2m2 for continuous culturing of up to 5E+10 cells. Two cartridges can be operated per Duet pump. The molecular weight cutoff recommended for exosome production is 20 kD but if a more complete cell secretome is required, to include small proteins, then 5 kD cartridges are available.

Simple routine exosome collection process

A FiberCell Hollow Fibre Bioreactor provides a continuous production process enabling periodic harvesting of concentrated EVs in 3ml, 20ml or 60ml syringe volumes. No passaging is required. Users of this system avoid batch production in cumbersome monolayer culture set-ups and will make major savings on disposable plasticware. Click on the article below to learn more.

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

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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