Nov 12, 2015 hollow fibre membrane bioreactor system. The hollow fiber bioreactor and cell cocultivation. Hepatic hollow fiber bioreactors are considered a promising class of bioartificial liver assist device blad. Hollow fiber bioreactors consist of a bioreactor, an oxygenator, a flow path, and a recirculation pump. A micro hollow fiber bioreactor and method of use are provided for use in screening different cell lines and process conditions. For adherent cell types, hfbs offer an in vivolike microenvironment as each fibre replicates a blood capillary and the mass transfer rate across the wall is independent from the shear stresses. C3 has four hollow fiber bioreactor hfbrx systems to address a wide range of production applications. Its a timescalable production process thats ideal for generating significant amounts of very concentrated evsexosomes, mabs or recombinant proteins.
Hollow fibers with 10 kda molecular weight cutoffs are housed within a piece of silicone tubing. Improved oxygen delivery to hepatic hollow fiber bioreactors. Crosssection of hollow fibers in fibercell systems hollow fiber bioreactor. C3s hollow fiber bioreactor systems are a solution to both goals. Hollow fiber membrane life in membrane bioreactors mbr. Fibercell systems hollow fibre bioreactor for high density. The contributions of several bioreactor designs and culture parameters, including the bioreactor geometrical parameters e. Hollow fiber membrane bioreactors have shown their advantages in various applications outlined in this chapter and beyond. Cd63 and alix were greatly enriched from the bioreactor compared to flask culture. Hollow fibers are commonly used substrates for specialized bioreactor systems, with the ability of some hollow fiber cartridges to culture billions of anchoragedependent cells within a relatively low bioreactor volume. Hollow fiber bioreactors are the focus of scientific research aiming to mimic physiological vascular networks and engineer organs and tissues in vitro.
Hollow fiber bioreactors introduction achieving high cell densities and reducing the costs to produce biologics are desirable but not always simple or inexpensive goals. Advances in fiber construction and methods for oxygenation have the potential to bring the advantages of hollow fiber cell culture to biomanufacturing. The hollowfiber bioreactor provides a defined and stable geometry, which allows for the control and management of the cell culture environment. The hollow fiber bioreactor system delivers media to the cells in a manner akin to the delivery of blood through the capillary networks in vivo. Combined ultrafiltrationtransduction in a hollowfiber bioreactor facilitates retrovirusmediated gene transfer. Hollow fiber membranes are ubiquitously used in industrial separations, especially the filtration of drinking water. It is expected that this paper can provide improved understanding on the key design parameters of the submerged hollow.
The reason for this lies in the interesting features of this bioreactor type, including excellent mass transport properties. Jul 27, 2014 hollow fibre membrane bioreactors hfb provide a novel approach towards tissue engineering applications in the field of regenerative medicine. Pdf hollow fibre membrane bioreactors for tissue engineering. Unlimited viewing of the articlechapter pdf and any associated supplements and figures. Production of exosomes in a hollow fiber bioreactor. Although the concept has been proved, detailed research and development are needed to translate the laboratory research into clinical applications.
Supported pvdf capillary fiber highly chlorine tolerant membrane high fluxlow pressure operation greatly reduced footprint as compared to conventional treatment hollow fiber membranes for membrane bioreactor applications in wastewater treatment worldwide sales and field service team proven engineering history membrane system design support. Pdf optimizing t cell expansion in a hollowfiber bioreactor. Cell and medium compartments of the bioreactor are separated by a semipermeable membrane, and oxygenation of the cell compartment is accomplished using an oxygenation membrane. The performance of a hollowfiber membrane bioreactor hfbr molecular weight cutoff 30 kd, fiber surface area 2050 cm2 containing a culture of hybridoma. Kdbio distributor for the fibercell systems hollow fiber. Automation and control system for fluid dynamic stability. Galactosylated polyvinylidene difluoride hollow fiber. The fibercell hollow fiber bioreactor continuous 3d culture at high density to. Hollow fiber bioreactors for in vivolike mammalian tissue. The hollowfiber membrane systems have been used as bioreactors years ago due to the advantages that present, such as. Use of a hollow fiber bioreactor to collect extracellular. Projects to generate grams of monoclonal antibody have been successfully managed with this system, for example, in cell. The ends of the fiber bundle are potted to form tubesheets slicing the tubesheets provides access to the fiber interior or lumens. It is a wellestablished system for the production of secreted proteins from hybridoma, chinese hamster ovary cells cho, human embryonic kidney hek 293 and other.
Each bioreactor contains thousands of hollow fibers that simulate living capillary systems. A dual hollow fiber bioreactor, consisting of an outer silicone membrane for oxygen supply and an inner polyamide membrane for substrate permeation, was used as an immobilized enzyme reactor to carry out enzymatic glucose oxidation. The system is designed to fit onto the shelf of a standard co 2 incubator and is simple enough to be setup by any competent cell biologist with a good understanding of aseptic technique. Immobilized mammalian cell cultivation in hollow fiber bioreactors. We have analyzed the design and operation of hollow fiber bioreactors for cell culture on the basis of cell growth. Interestingly, the galactosylated pvdf hollow fiber shows enhanced serum albumin diffusion across the membrane. Primary adherent cell types can be expanded in the quantum cell expansion system quantum system, an automated platform that utilizes a hollowfiber bioreactor. Hollow fiber membrane modules are used for a wide range of applications including dialysis 1, filtration 2, contactors 3, and gas separations 4. Hollow fiber membrane an overview sciencedirect topics. Filtration principles, operation, and membrane fouling ultrafiltration hollow fiber membrane bioreactor mbr treating oil refinery wastewater. The fibercell system a single fibercell systems hollow fiber cartridge is equivalent to harvesting 20200 roller bottles daily. The bioreactor includes the use of an oxygen permeable e.
This problem limits hepatocyte functional capabilities. Highdensity cell growth is shown in extracapillary space. Diffusion and inhibition processes in a hollowfiber membrane. The hollow fiber bioreactor disposable set is a highly efficient bioreactor configuration suitable for small scale biomass or protein production. Hollow fiber bioreactors for mammalian cell culture nature. A hollow fiber bioreactor is a 3 dimensional cell culturing system based on hollow fibers, which are small, semipermeable capillary membranes arranged in parallel array with a typical molecular weight cutoff mwco range of 1030 kda. Indeed, hollow fiber bioreactors allow limitations to be overcome in nutrient transport by diffusion, which is often an obstacle to engineer sizable constructs in vitro.
Largescale production of lentiviral vector in a closed. This manuscript describes in detail the setup and operation of a hollow fiber bioreactor system for the in vivolike culture of mammalian cells. Hollow fiber bioreactors have been in use for more than 30 years for the production of secreted proteins and antibodies from mammalian cells. A fibercell systems hollow fibre bioreactor enables the continuous culture of large numbers of cells at high density. Sa fiber count fiber length atf rate cycle time residence time process bioreactor m2 nominal cm lpm sec sec l lhr lpm lday wvday mlmin sec volume filtration rate scaleup parameters flow per fiber shear flux atf to filtration1 lmh ratio atf2h 0. Contrary to popular belief, it is possible to harvest large numbers of cells from a hollow fiber.
May 26, 2016 the hollow fiber bioreactor system delivers media to the cells in a manner akin to the delivery of blood through the capillary networks in vivo. Application of submerged hollow fiber membrane in membrane bioreactors. John j s cadwell, president and ceo, fibercell systems inc. Setting up a fibercell systems hollow fiber bioreactor. The pdf file you selected should load here if your web browser has a pdf reader plugin installed for example, a recent version of adobe acrobat reader if you would like more information about how to print, save, and work with pdfs, highwire press provides a helpful frequently asked questions about pdfs alternatively, you can download the pdf file directly to your computer, from where it.
The hek293 bioreactor culture yielded an equivalent number of exosomes per harvest as 70 t225 flasks. Pioneered hollow fiber bioreactor platform spunoff as cell culture company january 2016 33,000 sq. Automation and control system for fluid dynamic stability in. This new layout has shown good results in terms of hepatic functions i. Us10577582b2 method of reseeding adherent cells grown in a. Typically, the modules consist of a bundle of randomly packed hollow fibers. Freshly isolated rat hepatocytes were seeded and cultured in the extralumenal space of the hollow fiber cartridge for 18 days in a continuously circulated system.
Pdf hollow fiber bioreactors are the focus of scientific research aiming to mimic physiological vascular networks and engineer. The hollow fiber bioreactor hfbr is a highdensity continuousperfusion culture system consisting of a cartridge containing thousands of semipermeable hollow fibers in a parallel array within a tubular housing. Hollow fibre membrane bioreactors hfb provide a novel approach towards tissue engineering applications in the field of regenerative medicine. Unfortunately, the development of this type of device is currently hindered by oxygen limited transport to cultured hepatocytes, due to the low solubility of oxygen in aqueous media. This invention is directed to methods of directly reseeding harvested adherent cells grown in a hollow fiber bioreactor. One of the often considered methods for cultivation is by immobilization of the cells in hollow fiber bioreactors hfbrs. Cellmax cellmax duo hollow fiber cell culture system. The closed environment of the bioreactor remains intact during the cell culture process, eliminating open events and handson. From the oxygenator the medium flows into the hollow fiber bioreactor e through the lumen of each hollow fiber within the hollow fiber bundle where it perfuses the cells growing on the outside of the fibers with fresh nutrients and oxygen and removes metabolic waste products. Optimizing t cell expansion in a hollowfiber bioreactor. Hollow fiber bioreactor technology for tissue engineering applications article pdf available in the international journal of artificial organs 391 february 2016 with 702 reads. Hollow fiber bioreactor technology for tissue engineering applications.
The hollow fiber bioreactor for the reasons outlined above, there is currently tremendous interest in 3dimensional cell culture systems. For adherent cell types, hfbs offer an in vivolike microenvironment as each fibre replicates a blood capillary and the mass transfer rate across the wall is independent from the shear stresses experienced by the cell. Design and operating criteria for hollow fiber bioreactors. The hollow fiber bioreactor provides a defined and stable geometry, which allows for the control and management of the cell culture environment. Cells suspended in media are introduced into the space of the cartridge out side the fibers known as the exctracapillary space. Hollow fibers are commonly used substrates for specialized bioreactor systems, with the ability of some hollow fiber cartridges to culture billions of anchoragedependent cells within a relatively low download pdf. Hollow fiber bioreactors smallscale systems hf primer excellent alternative to tflasks, spinners, roller bottles, ascites, wave, celligen runs weeks to months yes uses singleuse disposables yes culture suspension or adherent cells yes bioreactor culture volume and size 80 ml,1.
A better way to grow cells single page pdf download. At the heart of every fibercell hollow fiber bioreactor system is the duet pulsatile pump. The potential application of hollow fiber bioreactors to. Jun 12, 2012 use of a hollow fiber bioreactor in stromal cellsuspension cell interactions. Instead, it will focus on suggestions that should be applicable to a. Hollow fiber bioreactors are the focus of scientific research aiming to mimic physiological. Hollow fiber is one of the most popular membranes used in industry. In order to more closely approximate in vivo cell growth conditions, richard knazek developed the hollow fiber bioreactor hfbr in 1972 1.
New methods of largescale vector production have therefore been contemplated, and one of the most prominent concepts is a closed system hollow fiber bioreactor. Jun 14, 20 demonstrating the first steps in setting up a hollow fiber bioreactor for preculture. This is integrated into a compact platform that supports one or two independent fibercell cartridge modules with associated media reservoirs. Conversion of bioreactors to continuous perfusion using.
The hfbr system is a highdensity, continuousperfusion culture system. The closed environment of the bioreactor remains intact during the cell culture process, eliminating open events and handson tasks associated with manual flaskbased culture. Perhaps the most rigorous application of hollow fiber systems for cell cocultivation is in the area of stromal cellsuspension cell interactions see figure 6. I chose a hollow fiber bioreactor with a low molecular weight cutoff mwco, 5kda because i was interested in collecting 100% of a larger, secreted mutant protein 35 kda in the ecs. Demonstrating the first steps in setting up a hollow fiber bioreactor for preculture. Glucose oxidation in a dual hollow fiber bioreactor with a. Filtration principles, operation, and membrane fouling ultrafiltration hollow fiber membrane bioreactor mbr treating oil refinery wastewater hollow fiber membrane modules hollow fiber membrane. Oxygen provision to hepatocytes and other types of cells maintained within the extracapillary space of a hollow fiber hf bioreactor is believed to be transport limited because of the low solubility of oxygen in aqueous media and the high cellular oxygen demand. It can be used for the cultivation of microbial plant or animal cells.
Pdf hollow fiber bioreactor technology for tissue engineering. Oxygen transport within a hepatic hollow fiber bioreactor. Also disclosed is a novel harvest media for use in directly reseeding adherent cells into a hollow fiber bioreactor. Report hollow fiber membrane life in membrane bioreactors mbr. Harvest large quantities of secreted protein with hollow.
Application of submerged hollow fiber membrane in membrane. Expansion and cellular characterization of primary human. Hollow fiber reactors reduce cost, scaleup concerns. This system can replace manual cell culture and produce cells that retain their phenotypes and functionality. Since there are a wide variety of different bioreactors being used for mammalian cell culture, this document will not address specific bioreactor models. Fibercell systems hollow fibre bioreactor for high density cell culture.
Hollow fiber bioreactors for mammalian cell culture. These hollow fiber membranes are often bundled and housed within tubular polycarbonate shells to create hollow fiber bioreactor cartridges. The hollow fiber usually has a membrane coated on the outside of a porous fiber support as shown in fig. Combined ultrafiltrationtransduction in a hollowfiber bioreactor facilitates retrovirusmediated gene. I first primed the hollow fiber bioreactor cartridge and confirmed the sterility with pbs for a minimum of 24 hours prior to preculturing the system with 2 l of. Us10577582b2 method of reseeding adherent cells grown in. The system conveniently fits inside a standard co2 incubator. The terumo quantum cell expansion system is a hollow fiber bioreactor designed to expand cells to the 10 9 range in a manageable and compact culture vessel. The use of cell cultures within a hollow fiber bioreactor could address many of these limitations and produce a continuous source of highly concentrated evs without contamination from serum evs, and that are suitable for downstream applications. Hollow fibre membrane bioreactors for tissue engineering. Hf bioreactors were first demonstrated by wolf and munkelt wolf and munkelt, 1975, who cultured hepatocytes in the extracapillary space ecs of a bioreactor derivative of an original. Hollow fiber bioreactor technology for tissue engineering. Jan, 2006 interestingly, the galactosylated pvdf hollow fiber shows enhanced serum albumin diffusion across the membrane. Fundamental research of factors affecting cell growth in hollow.