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Regarding suitability of Eppendorf bioprocess equipment in GMP-regulated applications, please reach out to your Eppendorf sales representative.
Stem Cell Bioprocessing
Stem cells are unique in that they possess the capacity for self-renewal and to develop into various cell types. This ability makes them extremely useful for biotechnological applications such as cell therapy , drug discovery, and the creation of modern food. Research and development in these application areas require large numbers of high-quality cells. However, establishing and growing highly sensitive cell cultures outside of their natural physiological environment is a labor and time intensive process. It is essential that the ideal conditions for cell growth can be produced and controlled, and that this system is scalable to generate the large quantity of cells required for cellular applications. Stirred-tank bioreactors facilitate efficient cell expansion through tightly controlled regulation of key parameters and the use of automation.
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Webinar: Breaking boundaries in bioprocess development
Join experts from Hannover Medical School, Pluri Inc., and Eppendorf and find out more about unleashing the power of stirred-tank bioreactors for cell and gene therapies!
New to stem cell bioprocessing? Find your answers below.
What are bioreactors?
The success of stem cell bioprocessing relies on robust and reproducible culture conditions and processes. Bioreactors are vessels used to culture a broad range of cell types in precisely controlled conditions to provide optimal productivity, efficiency, and product quality. Bioreactors come in a range of sizes from milliliters to hundreds of liters. They can either be single-use or made of glass or stainless steel.
For more information, check out our introduction to bioprocessing, where we ask – what is a bioreactor?
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What are the advantages of bioreactors over cell culture flasks?
Although widely used in stem cell research and production due to their ease of use and affordability, cell culture flasks are limited in terms of control and scalability. Bioreactors offer many benefits, such as:
- Better reproducibility: In bioreactors you can monitor critical culture parameters including pH , Dissolved Oxygen (DO) , temperature and nutrient feeding and automatically adjust them by bioprocess monitoring and control software. Like this, culture conditions and as a result culture outcome can be better reproduced.
- Simpler scale-up: Many stem cell applications in cell therapy development, drug discovery, and lab-grown meat require high cell numbers. When using a bioreactor one can increase the size of the culture vessel to many liters and even more, unlike cell culture flasks where the number of vessels must be increased after a certain point. Using one big vessel instead of many small ones saves space and manual labor, thus simplifying the process of increasing the number of cultured cells.
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Gain a more thorough understanding of the benefits of using a bioreactor with our white paper .
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How can I cultivate adherent stem cells in bioreactors?
The majority of stem cells are adherent cells and require a support matrix to grow. In 2D cultures, this means growing stem cells in dishes or flasks, however, these systems are often challenging to scale-up. Fortunately, there are many available options to grow adherent cells in bioreactors including:
- Cell-only aggregates: This technique involves allowing cells to adhere to one another and form an aggregate. This technique is often used for the cultivation of induced pluripotent stem cells (iPSCs) in bioreactors.
- Microcarriers: Microcarriers are spherical beads, which are added to the culture medium and serve as a surface for adherent cells such as mesenchymal stem cells (MSCs) to attach and grow. Microcarriers are made of a variety of materials. They can have specific coatings to enhance cell attachment, yield and viability.
- Fibra-Cel® Disks: Disks made of polyester and polypropylene provide a growth support for adherent cells in both packed-bed bioreactors and in suspension. Due to their three-dimensionality and fibrous material, Fibra-Cel Disks allow for easy cell attachment, a high surface-to-volume-ratio, and provide a low-shear environment. This often results in increased total biomass compared to many microcarriers.
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Discover the advantages of different matrix systems for adherent cells in our white paper .
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Can I also differentiate my stem cells in a bioreactor?
Efficient differentiation of stem cells into the desired cell type is critical for cell therapy applications, disease modeling, and the production of cultured meat. Stem cell differentiation is influenced by the culture conditions, such as the medium composition, physiochemical environment, cell-cell and cell-matrix interactions, and more. Bioreactors are an excellent platform that allow you to control, optimize, and reproduce the precise conditions required for specific cell differentiation. Stem cells have been successfully cultured and differentiated into different cell types using Eppendorf bioreactors. Please have a look at the documents below for some examples:
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Can I cultivate iPSCs as well as MSCs in bioreactors?
Both induced pluripotent stem cells (iPSCs) and mesenchymal stem cells (MSCs) can be grown in bioreactors, however it's likely that alternative cultivation methods will be required since iPSCs are often grown as cell aggregates while MSCs are often grown utilizing microcarriers. Find some examples below:
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How can I switch from stem cell culture in flasks and dishes to bioreactors?
The first step in optimizing your stem cell bioprocess is to switch from 2D to 3D processes for improved scalability and control. Here we offer some tips and tricks for process transfer to bioreactors and share the experiences of experts.
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Infographic: Stem cell culture in bioreactors
Download our infographic for some tips to help you transfer your stem cell culture from dishes and flasks to bioreactors.
Interview: Increasing hiPSC yields
Robert Zweigerdt fromHannover Medical Schoolexplains how his groupconverted stem cell culture to bioreactors and which process optimization measures were crucial to increase the cell yield.
Panel discussion: Preparing your stem cell cultivation for commercialization
At what point in yourdevelopment should youconsider which cultivation method? How to switch from static to stirred-tank cultivation? Listen to experts from the Cell and Gene Therapy Catapult and Eppendorf to discover answers to these and more questions.
Find more expert advice on stem cell bioprocessing here .
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Stem cell expertise at Eppendorf
Eppendorf has a long, established history working with stem cell processes in 3D cultures. Since 2012, Eppendorf bioreactor systems have been used to grow stem cell cultures in stirred tank bioreactors, presenting data towards scalable mass expansion of hiPSCs . Since then Eppendorf further developed its bioreactor design to suit the special needs of stem cells, for example as part of the EU Horizon 2020 research consortium Technobeat (Tools and TECHNOlogies for Breaktrough in hEArtTherapies).
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Choose the right bioreactor system
Bioreactor control stations
The DASbox® Mini Bioreactor System is widely used for stem cell applications. It has a small working volume of 100 mL to 250 mL and is therefore well suited to get started. If even higher cell numbers are required, process scale up to other systems, like the BioFlo® 320 or SciVario® twin is possible.
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Single-use bioreactors
The small and bench-scale Eppendorf bioreactor systems are compatible with BioBLU® Single-Use Bioreactors . Harboring a special 8-blade impeller, the BioBLU® 0.3sc was optimized for gentle mixing and cell aggregate formation.
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Our bioprocess specialists are happy to provide further support. Please contact us to discuss the needs of your individual application.
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Stem cell bioprocessing for the experts: Optimize your stem cell culture in bioreactors
How can I increase the yield of my stem cell culture in bioreactors?
Increasing cell density is a vital component of research in stem cell bioprocessing. Bioreactors are suitable tools, as they allow for monitoring and controlling the cell growth environment. Furthermore, they facilitate nutrient addition and waste product removal. One important consideration when optimizing your stem cell bioprocess is therefore to consider which operation mode is most suitable for your cells. A number of techniques exist including batch, fed-batch, continuous, and perfusion cultures and each has its own benefits and drawbacks.
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Find out, how the group of Robert Zweigerdt at MH Hannover increased hiPSC yield tenfold to 35 million hiPSCs per mL by optimizing process parameters and feeding strategy.
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How can I control the critical process parameters in my bioreactor?
Bioprocess monitoring and control
To obtain robust processes and cells of consistent, high quality it is indispensable to understand, which process parameters influence the cells’ characteristics. Learn, how the integration of sensors and control software into bioreactors enables real-time monitoring to gain a better understanding of the process and to automate process control.
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Learn from biologics bioprocessing
Cell culture in stirred-tank bioreactors to produce recombinant proteins is well established and the methods for bioprocess development are sophisticated. In stem cell bioprocessing the cell itself is the product and not a protein. What can the stem cell sector still learn from the bioprocessing of biologics? Read the interview with our stem cell expert Philipp Nold.
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Where can I find information about the cultivation of MSCs in bioreactors?
MSCs are multipotent cells derived from the bone marrow, adipose tissue, cord blood or other tissues related to the embryonic mesoderm. MSCs hold great potential for both cell-based therapies and for the creation of novel food. However, both of these industrial applications require robust manufacturing processes to increase the consistency and scalability of MSC production.
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Read our application notes for some guidelines on how to culture MSCs in bioreactors
Bone-marrow derived MSCs
PDF 0.9 MB
Adipose-derived MSCs
PDF 0.5 MB
iPSC-derived MSCs
PDF 0.3 MB
Where can I find information about the cultivation of iPSCs in bioreactors?
Robust large-scale hiPSC production is required to fulfill the large cell quantities needed in cell therapy development. Bioreactors provide a controllable culture environment and offer a scalable platform for culturing pluripotent stem cells.
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Read our application notes for some guidelines on how to culture hiPSCs in bioreactors
Reproducible production of hiPSC-derived neurospheres
PDF 1.9 MB
Reproducible production of hiPSC-derived cardiomyocytes
PDF 0.6 MB
How can I produce stem cell-derived exosomes in bioreactors?
Stem cell-derived exosomes are currently of great interest as cell-free therapeutic tools due to their strong diagnostic and therapeutical potential in various diseases models. To explore the use of exosomes in preclinical research, large amounts need to be produced.
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Join the stem cell community!
Organized for the first time in 2017, the Stem Cell Community Day (SCCD) connects researchers from academia and industry across the world, to discuss recent advances and new technologies in the field of stem cell production. With a special focus on the controlled cultivation of stem cells in stirred-tank bioreactors, the SCCD is an optimal place to exchange knowledge and share expertise with scientists around the world.
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