The principle of ‘Tide Motion’ in CelCradle contains the packed bed (BioNOC™ II) and the media reservoir in the same bottle. As the bellows are pushed up, the cells attached to the carriers will be submerged to the culture media, whereas cells are taking in nutrition. As the bellows are release, the cells are exposed to air in the environment taking in aeration.
CelCradle® bottle caps can be switched with a non-vented white cap for inoculation phase and a vented blue cap during cultivation phase. Culture medium can be drawn out through pipetting a small amount and measuring its pH through a pH meter, glucose through GlucCell™ or a bioanalyzer. Adherent cells attached to the macroporous carriers can be monitored through cell sampling. The bottle is opened inside a biosafety cabinet and using a sterile forceps, carriers can be taken out for cell staining and monitoring.
Usually, 2 to 3 pieces of carriers are required for every cell count procedure. However, it is suggested to do adherent cell monitoring thrice and stain cells with a dye to observe and count the cells. It is usually more difficult to detach bigger cells so make sure that cells are harvested using an optimized enzyme, accompanied with a mechanical agitation.
CelCradle® single use bioreactor is equipped with a CelFeeder™ for automated recirculation process during culture. The CelFeeder system is designed to simplify culture medium replacement through continuous harvest and feed. The daily feeding volume and the feeding frequency can be controlled during this process.
Using the CelCradle® 500AP bottle, tubings from the media reservoir bottle will be connected to its feed and harvest port. The media reservoir bottle shall contain fresh culture media in 1 L or 2 L volume and its cap to be replaced with a connection set. This connection set have replacement tubings and pump head for setting up the recirculation process mode.
Parameters such as cycles/day and volume can be set using this unit.
Change the bottle caps with the connection set provided in the CelCradle continuous complete system.
CelCradle® 500AP bottle is equipped with the feed and harvest port for connecting to the medium reservoir tubings.
Seeding adherent cells in CelCradle® bottles involved an upright and reverse method. In upright seeding, the carriers are pre-wetted with the culture medium to prime it. Prepared seed cells are pipetted into the disposable CelCradle bottle, topping the media to reach 500 mL, and set into the CelCradle® Stage. After 1-3 hours of incubation, the cell attachment efficiency will be calculated.
For reversed seeding, prepared seed cells are pipetted evenly on the carriers and the bottle will be capped of with a non-vented white cap, instead of the blue vented cap. The bottle will be inverted and placed inside the incubator, allowing cells to attach for 3 hours. One the first hour, while still inverted, the bottle is swirled every 15 minutes.
The CelCradle 500A and CelCradle 500AP disposable bottles are suitable for production of both secreted and non-secreted products. These two singe use bottles were designed to enhance the cell attachment efficiency (for loosely adherent cells) using the reverse seeding method.
Using a strainer, secreted product expelled from the medium can be harvested continuously and purified further to become the final product.
The CelCradle bioreactor is mostly used in bench-scale suitable in research and development laboratories whereas TideXcell-002 can be applied in both academia and small pilot tests. The final cell densities in both systems are related proportionally. However, the productivity and cell growth rate depend on several factors.
Since CelCradle system does not have automated pH control and perfusion process mode, the cell culture takes slower pace than TideXcell-002. With linearity of scale-up, TideXcell-002 is better than CelCradle. The parameter setting in bench scale can be used for the pilot scale, but some modifications are still necessary. Compared with other commercially available system, however, very few parameters are required to input in the TideXcell system; thus, optimization is quite straightforward.
Culture medium replenishment is important as cells consume nutrients from it. It is rather difficult to maintain 10 day-VERO culture in the system without exchanging the culture medium. However, the culture temperature can be reduced below 10°C to achieve high cell density. Addition of supplements such as glucose and glutamine concentrate can also be done to maintain cell viability.
Be sure to control pH within optimized range (usually 7.0-7.4) through addition of NaHCO3. However, the osmolarity will increase and affect the cell viability if too much sodium bicarbonate is added.
There are two methods for users to observe the cell culture performance.
The mechanism of CelCradle enables a fast and complete intake of nutrients and oxygen in the culture medium to the macroporous carriers (matrix). All ingredients in the medium are properly mixed while the CO2 produced by glucose hydrolysis can be expelled efficiently along the run.
The CelCradle® system also offers high surface area for adherent cell growth.
The output of 1 CelCradle® bottle is 2-3 billion VERO cells and around 200 million cells for mesenchymal stem cells. The cell harvest varies depending on the cell type, enzyme used, age of the cells, and more.
The system’s cell harvesting procedure is easy. Learn more about the protocol here to achieve more than 90% cell harvest.
|Dimension||264 W x 359 L X 170 H mm (10.4 x 14 x 6.7 inches)|
|Weight||7.0 kg(15.4 lb)|
|Power||100~230 VOLTS AC, 50/60 Hz (Input); 12 VOLTS DC (Output_)|
0.25 to 2.0 mm/sec.
Step of 1.0 sec or 1.0 min.
0 to 99 min 59 sec
Step of 1.0 sec or 1.0 min.
|Driver Motor||DC stepping-motor|
20 ~ 42 °C, 0 ~ 90% relative humidity
(in a CO2 incubator)
|Mechanical Protection||Hi-low Optical Sensor|
|Transmission||Gear set (ration 1:1.2) and belt|
|Recommended CO2 incubator dimension||
CelCradle Complete: CCL-170L model:
505 x 535 x 633 mm(19.9" x 21.1" x 24.9") and
CelCradle Continuous SYstem Complete: CCL-240L model:
595 x 640 x 633 mm (23.4" x 25.2" x 24.9")
|Dimension||137 L x 226 W x 40 D mm (5.4 x 8.9 x 1.6 inches)|
|Weight||1.16 kg(2.6 lb)|
|Power||100-230 VOLTS AC 50/60 Hz (input): 12 VOLTS DC (Output_)|
|Environment||Room temperature (outside CO2 incubator)|
|Materials||Magnetic back plate, to hold the controller to the side of the incubator|
|Dimension||230 W x 131 D x 61 H mm, (5.2 x 9.1 x 2.4 inches)|
|Weight||0.645 kg (1.42 lb)|
|Power||85~240 Volts AC, 50/60 Hz (input); ~180 mA|
|Environment||Ambient to 45°C, ~95% Relative Humidity|
|Application/Bottle||Item Code||Secreted Protein, Viruses||Cell Harvest
(for non-screted, proteins, viruses or cells)
(for protein, extraction or reuse of carriers)
|CelCradle® 500A||1400003||Applicable, but not optimal||Best application||Best application|
|CelCradle® 500AP||1400004||Applicable, but not optimal||Best application||Best application|
VacciXcell has to date sold over 600 CelCradle Single Use Bioreactors and have a growing portfolio of clients. Due to non-disclosure agreements (NDA) only limited information may be provided of our clients.
Some of these clients are:
|Country||Cell Line||Virus Strain||Cell Density||Virus Titer|