3D culture in a hollow fibre bioreactor

Simulate in vivo conditions

In a hollow fibre bioreactor cells can grow in 3 dimensions around porous, semi-permeable, protein-retentive “capillaries” that carry recirculating media on the luminal side. These are conditions that resemble the in-vivo situation much more closely when compared either to flask or microcarrier culture (N.B. microcarriers are often claimed to provide a 3D culture method but microcarriers only allow cells to attach and grow on the outermost surface which limits the formation of multidirectional cell–cell interactions).

Hollow fibre bioreactor fits in a standard incubator for convenient cell culture scale-up

Growth of cells around porous, semi-permeable hollow fibres, perfused with media. Small molecules such as glucose, metabolites and antibiotics can easily cross the fibres.

Highest cell density

In a hollow fibre cartridge cells grow in a 3D space at very high cell densities, around 1E+8 cells/ml,  and such cultures can be maintained continuously for long periods. Typically cell division will be slower than in T-Flasks.

Culture-supportive secretome

3D cultured cells in a hollow fibre cartridge will generate a secretome and microenvironment enriched in cell-derived EVs, cytokines etc. thereby creating more in vivo like conditions. Such conditions support the long-term maintenance of the culture and can be exploited to generate in vitro tissue models. Objectives might include proteomic analysis or harvesting of target components such as exosomes, or perhaps the harvesting of a different cell type grown in co-culture (see below).

Collect enriched secretome

Highly enriched conditioned media can be periodically collected, e.g. for secretome analysis / proteomics or to obtain large amounts of concentrated extracellular vesicles. The culture of primary MSCs in the 20kDa FiberCell cartridge is a well-established application, for example. Protein- and EV-retentive 5Kd or 20Kd molecular weight cut-offs are available for polysulfone fibres. Special 0.1µm or 0.03µm microporous PVDF fibres are also available which may offer tighter protein matrix binding to enhance cell attachment.

Easily replace serum

Under high cell density bioreactor conditions that are naturally more supportive of growth it is usually possible to completely replace serum with a chemically-defined substitute called CDM-HD. This is a defined powder mix contains only low molecular weight components. A FiberCell bioreactor using CDM-HD media can further help the researcher to recapitulate in vivo growth conditions.

Co-culture

Successful co-culture tissue modelling has been achieved using the FiberCell bioreactor, for example to create a bone marrow model or to culture endothelial cells under flow conditions on the luminal side of 0.1µm pore size fibres. Article: 3-D in vitroModels using Hollow Fiber Bioreactors

Setting up for 3D culture in your lab

A complete FiberCell Hollow Fibre Bioreactor including media reservoirs will fit neatly on the shelf of a standard lab incubator, or inside a hypoxic chamber, for continuous culture of more than 1E+9 cells in a 20ml cartridge. Learn more about hollow fibre culture.

Hollow fibre bioreactor fits in a standard incubator for convenient cell culture scale-up

A FiberCell® Bioreactor, complete with media reservoirs, will easily fit inside a standard incubator. No special connections required.

3D Culture References:

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