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CelCradle® Benchtop bioreactor

Cradle for your Adherent Cells

CelCradle is a bench-scale bioreactor system that can support high adherent cell density culture. It is a single-use, cost effective packed-bed bioreactor system for adherent scale-up from laboratory scale to production scale. This disposable bioreactor fits inside a 6ft3 CO2 incubator (for batch culture) or 8.5ft3 CO2 incubator (for perfusion culture).

The ready-to-use bottles, as part of single-use manufacturing, helps eliminate validation costs as compared to multiple-use system when starting on your next cell culture run. The system components come pre-sterilized, pre-packed that speed up your research whether for vaccine, stem cell, or viral production applications.

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CelCradle<sup>®</sup>

Tide Motion in CelCradle®

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.

Tide Technology Features

Features

  • SS 316 L Carcass for easy cleaning

  • Compact and easy-to-install, small footprint benchtop design

  • Pre-sterilized, ready-to-use, disposable bottles

  • Conductive Environment

    - Culture cells with an environment of extremely low shear stress, foam-free, and no O2 limitation

  • Compatible with most serum-free media

  • Macroporous Carriers’ Surface

    - to grow most anchorage-dependent cells and allow easy harvest of whole cells, cell components, or secreted proteins

  • Provides large surface area of up to 15,000 cm2 per bottle for adherent cell growth

  • Capacity to produce hundreds of milligrams to several grams of secreted proteins or monoclonal antibodies per month in four bottles

  • Few parameters for easy optimization

  • Culture in batch, fed-batch, or recirculation modes

  • Scale-up through TideXcell pilot scale bioreactor or through directly multiplying number of bottles used

Applications

  • Mammalian and Insect Cell Culture
  • Monoclonal Antibody Production
  • Gene and Cell Therapy (autologous)
  • Viral Vector production
  • Pharmacokinetics Study
  • Vaccine Production
  • Drug Discovery
  • Protein Production
  • Proteome Research
Tide Technology Applications

CelCradle System Complete

Simple to operate and virtually cruve same CelCradle unit stage but utilizes CelCradle® 500 and 500A bottles
CelCradle<sup>®</sup> product 1 CelCradle<sup>®</sup>product 1 with label

CelCradle Continuous System Complete

Same CelCradle unit stage but utilizes CelCradle® 500P and 500AP for continuous process.
CelCradle<sup>®</sup> product 2 CelCradle<sup>®</sup> product 2 with label

Product Highlights

Cell Culture Monitoring

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.

Three Easy Steps to Monitor:

Pipette Pipette
Picking Picking
Stain Stain

Recirculation Process Mode

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.

Three Easy Steps to Monitor:

Set-up the CelFeeder Set-up the CelFeeder

Parameters such as cycles/day and volume can be set using this unit.

Prepare the Media Reservoir Bottles Prepare the Media Reservoir Bottles

Change the bottle caps with the connection set provided in the CelCradle continuous complete system.

Connect to the CelCradle Bottle Connect to the CelCradle Bottle

CelCradle® 500AP bottle is equipped with the feed and harvest port for connecting to the medium reservoir tubings.

Seeding Methods

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.

*Protocol varies on the type of cell line used

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.

Frequently Asked Questions

How Does It Work?

This disposable bioreactor only works for adherent culture. However, if the said suspension cells are loosely adherent and could attach in the macrocarriers, the secreted protein can be harvested for long periods with much lower cost than commercial suspension culture system.
There are several steps to detach cells from the carriers. One is through the use of a dissociation enzyme, either trypsin, collagenase, or more. The type of enzyme to be used for a certain process depends on the type of adherent cell being cultured. Secondly, mechanical agitation is required after enzyme treatment. This is to dislodge cells from the carriers to achieve more than 90% cell harvesting efficiency.
Two pieces of carriers are required for every cell counting procedure. However, it is suggested to perform the test thrice and then stain with CVD and multiply the averaged calculated cell number by 865 (amount of carriers/CelCradle bottle). The result would be the total cell numbers. Before trypsinization, check the reference number to make sure that cells can be detached and achieved more than 90% harvesting efficiency.
Mechanical agitation through rotation or shaking the bottle is required. Alternatively, the clients can also use the automated harvesting system.

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.

Cell culture growth curve is very important to know and check the cell’s confluency. It is recommended to perform cell growth test and to monitor cell growth daily before it reaches its plateau phase. The maximum cell density will vary depending on the cell line.
The virus infection depends on how the viruses infects cells. If the viruses could effectively infect the cells, infection procedure can be done the same way in cell seeding.
For cell harvest, inverted method for enzyme incubation is best to use as it will only require less enzyme to detach cells, less buffer media, and has higher cell recovery rate.

There are two methods for users to observe the cell culture performance.

  1. Way 1: to measure the metabolite consumption in the cultured media. The pH and glucose are the basic parameters to be monitored during culture.
  2. Way 2: to take out carriers from the bottle and count the cells using a stain.
Commonly used adherent cell lines such as VERO, HEK 293, CHO, Mesenchymal Stem Cells, MDCK, MDBK, MRC-5, and more can be cultured in the CelCradle system.

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.

  • 1 CelCradle bottle =
    • 18-20 pcs. of Roller Bottles (850 cm2)
    • 4 pcs. of Spinner Flasks (500 mL)
    • 100 pcs. of Petri Dish (180 cm2)
    • 2 pcs of Cell Stacks (6,320 cm2)
    • 125 pcs of T-flasks (150 cm2)
There are approximately 860 pieces of BioNOC™ II in every CelCradle bottle. The deviation is less than 5%.
The CelCradle bottle comes pre-packed and pre-sterilized so it will be ready to use for culture. It is NOT recommended to use your own carriers as the system is standardized and optimized with BioNOC™ II.
Without changing the gene of the cells, it is very unlikely to promote the cell propagation while limiting critical nutrients. With the same quantity of medium in a certain period of time, CelCradle’s maximum output may not be different from those of other commercial products. However, CelCradle does present high cell density due to the cell growth pattern.

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.

The BioNOC™ II carriers in the CelCradle bioreactor are negatively charged and should be okay for coating factors such as Matrigel, fibronectin, and more. These additional coating factors enable cells to attach more in the macroporous carriers.
The bottle can be placed in a -80°C freezer directly for free-and-thaw process to extract cellular components.
The CelCradle® bottles are pre-sterilized using gamma irradiation process.
The bottle is a ready-to-use bottle, designed for ONE time use only and cannot be autoclaved. Reusing the CelCradle® bottle is not recommended.
The bottle is composed of 3 materials: the main bottle part is made of PET, the bellow part is made of EVA, and the blue parts are made of PP.
The bottle uses PTFE hydrophobic filter with 0.22 µm pore size.
Users can sample and take out macroporous carriers from the CelCradle bottles, harvesting and staining the cells for observation. The carriers are made of porous non-woven fabrics. The common staining solution includes hematoxylin, trypan blue, calcein green, and more.
CelCradle comes with a Certificate of Qualification (CoQ) and Certificate of Compliance (CoC), and the stage is CE certified and comes with a Certificate of Qualification (CoQ).

Product Specification

CelCradle® Stage Technical Specifications

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_)
Up-Down Rate 0.25 to 2.0 mm/sec.
Step of 1.0 sec or 1.0 min.
Delay Time 0 to 99 min 59 sec
Step of 1.0 sec or 1.0 min.
Driver Motor DC stepping-motor
Environment 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")

CelCradle® Controller Technical Specifications

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

CelFeeder™ Controller Technical Specifications

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)		 Carrier Harvest
(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
  • CelCradle 500
  • CelCradle 500AP
  • CelCradle 500P
Esco VacciXcell Product Guide
Esco VacciXcell Product Guide
CelCradle<sup>®</sup>
CelCradle®
Protocol for Generation of HEK293T Seed Train Using CelCradle - EN
Protocol for Generation of HEK293T Seed Train Using CelCradle - EN
Protocol for Culturing MSCs in CelCradle™ - EN
Protocol for Culturing MSCs in CelCradle™ - EN
Cultivation of hybridoma OKT 3 cells for MAb production
Cultivation of hybridoma OKT 3 cells for MAb production
Cultivation of MDCK cells
Cultivation of MDCK cells

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
USA CEF WEE 4 x107 109pfu/ml
Taiwan BHK-21 Pseudorabies 2.35x107 108.8 TCID50/ml
Taiwan HEK-293 Adenovirus 3.5 x107 108pfu/ml
Taiwan SF-9 Baculovirus 6x107 109pfu/ml
Esco Lifesciences Group