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Z®RP Technology
Cell Culture and Tissue Engineering with
Glenmills/Zellwerk Z®RP"Cells Working for You"
Frequently asked questions and answers concerning Z®RP Technology
What types of cells can I grow in a Z®RP BIOREACTOR?
All types of eukaryotic cells: mammalian cells, human cells, insect cells, plant cells, yeast,
filamentous fungi, bacteria, adherent growing cells, suspension cells, cell lines, primary cells,
stem cells, progenitor cells, embryonic stem cells can be seeded and expanded in Z®RP
Bioreactor®.
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What are the main characteristics of cell culturing in a Z®RP BIOREACTOR compared to
stirred bioreactors and other systems?
Z®RP Bioreactor come with a variable rotating bed and work in perfusion
mode where the cell medium pours over the cells on a continuous basis by virtue of the rotation.
Cells grow attached to the bed in a three-dimensional fashion while embedded in their own extra
cellular matrix. Gentle moving of attached cells alternately through medium and overlay
guarantees excellent supply with nutrients and oxygen (or other media) without being subjected
to shear stress.
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What is a rotating bed?
The term "bed" refers to various types of carriers or scaffolds that can be applied in a Z®RP
Bioreactor system. The standard rotating bed comprises of Sponceram® carrier discs manufactured from proprietary ceramic materials with adjustable adhesiveness and porosity. Many other bed materials are available as well as holding fixtures for experimental scaffold materials. Customized scaffolds and carriers of all kinds can be realized.
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Will I be able to detach my cells from Sponceram® carriers?
Yes, detaching cells is achieved by incubating the attached cells with suitable enzymes (trypsine,
collagenases e. g.) and/or mild detergents in conjunction with enhanced rotating speed. Detaching
protocols vary depending on total cell mass, rinsing procedures, carrier or scaffold type and cell
type. Refer to these application notes in PDF format for more details on removing the cells from the scaffold.
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How many cells can I harvest from a Z®RP BIOREACTOR?
In a Z®RP 500ml vessel equipped with full number of carriers 1 to 2 x 10^10 can be harvested typically. In a 5000 ml vessel it would be 1 to 2 x 10^11 cells typically. The latter is about the number of cells of a human liver.
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Can I work with small cell numbers in a Z®RP system?
A Z®RP system allows you to work with cell numbers as low as 1 x 10^4 cells. You can adjust the
culturing volume by using our small disposable vessel types or use spacers in a 500 ml vessel or
order a customized rotating bed for your application. Even with the smallest vessel you do have
the full functionality of the Z®RP TECHNOLOGY with respect to process control and
reproducibility.
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Can I grow and harvest stem cells in a Z®RP BIOREACTOR?
You can culture and expand all types of stem cells in a Z®RP system. Harvesting procedures
depend on cell type, carrier type and media used.
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Will stem cells differentiate when cultured in a ZRP system?
Depending on carrier type and medium used you are able to optimize your culturing protocol for
minimum or maximum differentiation. Induction of specific differentiation is possible by
addition of growth factors, signal transduction factors, scaffold properties and culturing
conditions. Co-culturing with other cell types e. g. feeder cells - is a third option to enhance
growth and control differentiation.
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How long can I run a Z®RP BIOREACTOR?
A Z®RP system loaded with cells can be run indefinitely depending on the type of cells. Longest
runs to date with mammalian production cell lines lasted almost one year and were intentional
aborted.
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Is the GMP BREEDER necessary for operation of a Z®RP BIOREACTOR?
Yes, the GMP Breeder® provides gentle and precise air heating for the
bioreactor vessel and its controls. A GMP Breeder avoids hot spots common
with water jacketed vessels. Moreover, it provides a sterile working environment for many kinds
of manipulations such as sample taking and harvesting.
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Can I use my own culturing vessels in a Z®RP system?
In some cases it is possible to adopt your own culturing vessel to the system. Additional costs
depend on features that have to be included into the system for proper usage of your culturing
vessel.
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Can I use my own scaffold/carrier in a Z®RP BIOREACTOR vessel?
Yes, customized scaffold holders can be provided. You can use all kinds of scaffolds as long as
they are fitting into the vessel. For large tissue engineering constructs you may use the 5000 ml
vessel.
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Are there disposable vessels for a Z®RP BIOREACTOR available?
Disposable vessels will be available in summer 2007.
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What types of disposable vessels will be available?
One chamber vessels for expanding small, medium or large numbers of cells with disposable pH-
and PO2-sensors, two chamber vessels for culturing stem cells under the influence of feeder cells,
customized variable scaffold vessels for culturing your cells on your scaffold
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Is the Z® RP TECHNOLOGY suitable for co-culturing of cells?
You can inoculate more than one type of cells in one bioreactor run through the inoculation port
of the system. Rotating beds with membrane separated chambers are also available for co-
culturing applications.
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How do I harvest my medium from the vessel?
The perfusion system of a Z®RP system allows you to harvest the used medium in any interval
you like. Just change the appropriate media collecting container in the sterile GMP Breeder environment.
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How do I change the medium source during a long lasting perfusion run?
The GMP Breeder allows for sterile exchange of media containers during
long lasting perfusion runs, if all necessary precautions for sterile work are taken.
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Can I use the Sponceram® carriers more than once?
No, Sponceram® carriers were designed for single us only.
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Is the Z®RP system compliant with GMP/GTP/GCP requirements?
Yes. If you use your Z®RP system in regulated areas and need equipment qualification
(IQ/OQ/PQ), we will provide full documentation by supplying reports on installation, operational
and performance qualification on request. Additional costs will apply.
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Do I need to take precautions to ensure sterility in the GMP BREEDER?
Yes. Even though a GMP Breeder is equipped with efficient UV-light
sterilization and HEPA filter and its design allows for laminar flow bench type operation you
need to make sure, that no contaminants are carried into the GMP Breeder.
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Can I use the system for tissue engineering applications?
Yes, a Z®RP system can be equipped with many types of cell carriers and scaffolds to produce a variety of autologous or allogenous products. A Z®RP system is suitable to be applied in standard clinical environments.
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Which process parameters are controlled?
Oxygen partial pressure, pH, GMP Breeder temperature, vessel temperature, circulation volume, media feed, base pump speed, CO2-provision, N2, O2, air, sterilisation cycle, rotational speed Bioreactor.
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Does the Z®RP CONTROL SYSTEM provide data logging?
Yes, your process data will be written on the personal computer linked to the Z®RP Control System. For safety reasons a significant time period worth of data is intermediately logged in the control itself.
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Can I take samples from the medium during operation?
Yes, medium samples may be drawn from a septum port in the circulation tube located within the
GMP Breeder.
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Can I remove cells from the carriers during operation?
Some disposable vessel types will allow the removal of cells from carriers during operation by
syringe or spatula action through a septum port.
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Are process data available on a PC?
Yes, all process data can be transferred to a PC using a common file format. A Z®RP system comes with control access and visualisation software as well as USB cable for connection to the PC.
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Is it possible to include a tangential-flow filter in the media recovery path to concentrate protein product?
Yes, filtering devices could be installed in the circulation tubing as well as in the run-off tube. The latter solution is preferred in order to reduce the risk of contamination
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Would it be possible for me to look at brochures or manuals on the system? Especially on the controller and the sensor systems? Exactly what parameters can be controlled?
More detailed technical brochures contain information on the GMP Breeder, the Control Unit and the sensor system are available upon request.
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What is necessary for running the system that is NOT included?
Air, carbon dioxide and power must be supplied. Depending on the cell type oxygen might be needed too.
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Can more than one reactor be installed in the breeder?
No.
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Can the controller run more than one reactor/breeder unit simultaneously?
No. However, a 10-fold up-scaled bioreactor (5 liter reactor vessel) is available and can be driven and controlled within the Z RP equipment. A process can be developed using the Z RP 500 bioreactor and then easily transferred onto the Z RP 5000 bioreactor.
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Can we get the test kit to make sure our cells will attach and grow on the Sponceram substrate?
Yes, even though cultivation results are much better under dynamic conditions.
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Is there any literature on the production of recombinant proteins using the system? Are there any sample protocols for the production of recombinant proteins? (I am interested in including in the cost/benefit analysis such things as media consumption, operator time, volume & concentration of product, etc.) In fact, do you have an estimate of the operating cost?
Since recombinant protein production is dependent on the cell line, media used and other factors cost/benefit analysis must be done on a case-to-case level. For some highly glycosylated secreted proteins we found Z®-RP cost of ownership very low when compared to normally used perfusion processes.
The reasons are:
- 3D culturing allows use of simple media.
- Much less supplementation is needed (etc. insulin, other growth factors) due to the fact that cell-typical extracellular matrix binds growth factors.
- Product yields in the media are higher compared to other cultivation procedures (roller bottles, stirred fermentors, hollow fiber reactors etc.).
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How long is the training class?
2 days.
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What kind of warranty and post-purchase support is provided?
12 months of warranty on device parts.
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How quickly can the system be delivered once it is ordered? Is there a long delay while a system is built, or is it a matter of just packing one up and shipping it? Who installs it? Who fixes it?
The system will be delivered approximately 4 weeks after getting an order. Usually the 2 days training is sufficient for the user to install and become familiar with the system. Furthermore, a comprehensive description of the system gives answers to most questions. Additional support will quickly be given by Glen Mills and/or Zellwerk per email or phone. The same is true for fixing problems.
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