WELCOME TO 

THE CELL CULTURE REVOLUTION

Introducing the Petaka G3

Transforming the art of cell culture into scientific engineering.

The Celartia cell culture system, based on Petaka G3, applies scientific reasoning and sound engineering to transform the art of cell culture into a science of standardized results.

.Our clients come to us interested in the Petaka because it mimics native-like conditions and provides more biologically relevant data. In addition it enables live cell room temperature shipping for up to 15 days.

It is essentially in itself a mini incubator. All you need is a heat source.

Key Features


- Addresses the age-old issue of hyperoxia.
- Cells are usually exposed to 3-5x the amount of oxygen in traditional flasks/incubators.
- The Petaka reduces controls O2/CO2 levels within the flask to mimic native like conditions.
- Culture more cells in each Petaka and add 1-2 days to media changes
- Slim Design - can provide up to 15x more cell culture surface area per incubator capacity
- Can be flash-frozen in vapor-phase nitrogen storage for long term storage
- Can do long term live cell benchtop microscopy studies - just add heat source
- Elimination of the water tray in the incubator (it’s a closed system)
- Reduction in plastic waste by 67%
- Individually barcoded for easy tracking 

Petaka's Key Features in Detail

Minimum Container Volume for the Maximum Number of Cells

For scale up cultures, Petaka offers important advantages.
Using Petaka enables you to maximize the efficiency of your incubator.
By minimizing the amount of air space in Petaka and simultaneously offering 150 cm2 surface area for cell attachment per unit, it offers up to 10 times more cell yield per incubator than T75-Flask or T160-Flask.
Moreover, whilst consuming proportionally less media per cm2 of surface saves media and growth factors per million cell yield.

Ship Live Cells in Petaka G3 Worldwide, Without Cryopreservation or Dry Ice

Up to 14-days at room temperature. Eliminate cryo-freezing and cold chain logistics for most applications. 

With the Petaka you can transport live cells for long distances at normal ambient temperatures. For most cell lines, PetakaG3 does not need dry ice or refrigeration to transport living cells for periods of time between 1 and 14 days.

Its slim shape permits easy packaging and protection. Especially designed mailers are ideal for shipping one or two units.
Cells inside Petaka, at normal temperatures, remain in a dormant state for days, even weeks. Viability decreases progressively depending on the cell type. CHO-K1 cells, for example, can travel for 3 to 4 weeks maintaining 75% viability. Some tumor cells can travel for a month or more.

Minimized maneuvers = time saving 

Inside the Petaka itself, one is able to perform several protocol steps without removing the cells from the device. This means that by not having to transfer cells to several other instruments or devices, one is able to cut out several steps. Examples of such applications are cell washing, cell & supernatant separation, enzyme-free detachment & harvesting, DNA & RNA extraction, cell pelleting, cell storage and pre-preparation of ready to host Petakas. Not only does this save time, but also reduces plastic waste by 67%. 

You can also directly flash-freeze the Petaka into Vapour-phase Nitrogen storage, eliminating the need for additional plastic use. 

Different Petakas for Different Cell Types

Centrifuge, Concentrate, and Wash Cells Directly in the Petaka G3

Eliminate transfers and contamination risk.

Petaka G3 simplifies operations by performing two functions, cell growth and cell concentration in a single device. Centrifugation of Petaka using the Petaka rotor, allows concentrate the cells in the corner of the media channel facilitating cell recovery with minimal media. Or concentration of the cells in the opposite corner allowing media harvesting avoiding the cell pellet. As a result, cell culture becomes simpler, quicker & more versatile.

For All Cell Types

Practically every cell type has been successfully cultivated in the Petaka G3™, including attached, suspension, immortal, primary and transfected
cells, hybridomas, MSCs, ESCs, iPSCs, etc. The Petaka G3™ FLAT is untreated for suspension cell culture. The Petaka G3™ LOT is surface treated
for attached cell culture. The Petaka G3™ is not just another flask; it corrects the common error of cultivating cells in the harmful, hyperoxic condition typical with plates, dishes and flasks. Since most cell lines have been cultivated in hyperoxic conditions for decades, Petaka users may observe differences as the cells re-adapt to their correct, physiological environment. This response may manifest as slight morphological changes,
changes in doubling time, etc., but this is not a negative effect. It is actually corrective, and in the long-term your cells will be healthier and will better reflect conditions in the native tissues when switched to grown inside the Petaka G3™.

Highly protected against contamination

Microbial contamination is a severe problem in cell culture. Typical routes of microbial infection in cultures are the ambient air, when flasks are transferred from the hood to the incubator, the water bath, and the humid environment of the CO2 incubator which provides ideal growing conditions for many strains of bacteria.

Other routes of cell culture infection include: contact with non-sterile surfaces when performing cell culture manipulations, spillage on materials and the work surface, splash-back from pipetting or pouring cell suspensions and microscopic aerosols.

Petaka is a virtually hermetically closed system. Air access is only possible through a 0.2 micron pore filter. Neither lids nor caps are used to access the liquids. Petaka are injected through a silicon port that seals after the removal of the injection tip.
Sterilizing the injection tips and the silicon port minimizes the chances of contamination. In addition, Petaka has an 80 mm diffusion barrier between the port and the culture chamber. This exponentially decreases the chances of microbial progression through the port slit during long incubations.

Syringe-based liquid transfers through sterile silicone port; eliminates open transfers and com

Long-Term Benchtop Microscopy

Because the Petaka G3™ provides a unique, closed, gas-controlled culture environment without significant medium evaporation, it functions like a mini-incubator all by itself. Because of this, specialty cell culture and experimental protocols may be performed for long periods of time outside of the standard CO2 incubator environment. This is impossible with plates, dishes, flasks or bags since medium will evaporate leading to loss of osmolarity, and the dissolved CO2 will rapidly be lost, resulting in loss of pH control and cell death.

The Petaka G3™ supports functions like time-lapse microscopy without any special (and expensive) microscope incubator equipment. All one needs to provide is warming for the Petaka G3™, everything else is automatically maintained by the device (see our longterm benchtop microscopy setup, right). At Celartia, we routinely use small heating films to maintain cultures at 37°C on a standard microscope platform to record multiday microscopy observations or to record pO2 in the medium on the benchtop.

Oxygen Concentration: Physioxia Within (down to 2% O2 inside, 21% O2 outside)

Petaka (Hypoxia Chamber) incubated in normal atmosphere (21% Oxygen) provides a gradually decreasing Dissolved Oxygen (DO) concentration in the media where the cells are cultured, always within the physiological limits (Physioxia) of living tissues cells (Graph 1).

Cell cultures begin with a DO concentration equivalent to that of arterial blood (Oxygen Partial Pressure 75 mmHg), high enough to promote exponential cell growth. As cells proliferate the concentration automatically reduces to those DO concentrations close to those levels that the cells receive in normal living mammalian tissues and in embryonic development, in which PO2 is close to 15 mmHg (see references), avoiding cell damage by ROS formation and facilitating cell differentiation.

Without doubt cultured cells require oxygen, but not in excess. When cells are cultured in media, in classic cell culture devices such as flasks and Petri dishes, cells are exposed to an excess concentration of DO. The open atmosphere contains about 20% oxygen. Therefore, the oxygen of the air dissolves in the media up to the limit of its solubility in water, depending on cell culture temperature, atmospheric pressure, and the degree of salinity of the media, resulting in DO concentrations that are far too high for normal cells integrating the living tissues. That is an artifact which can be the source of many misleading results and discrepancies between experimental setting and in vivo setting.

The CelartiaTM cell culture system automatically balances the partial pressure of the dissolved Oxygen in the media at 25 mmHg, reproducing the cell respiratory and growth conditions in the natural environment, that is, as if within a living being. Therefore, the PetakaTM bioreactors (patented) have been designed to provide semi-hermetic cell chambers with physiologic respiration, using a unique system of micro-tubular gas diffusion channels. With this system, the threshold of oxygen in the cellular environment is maintained at physiological levels, similar to the natural bodies of living creatures, producing a perfect environment for stem cells and all other mammalian cell types. Enabled to incubate the cells in an incubator with 21% Oxygen, the Celartia’s cell culture system reduces the Oxygen down to 5% or even as far as 2% under certain specific conditions depending on cell type and level of confluence, which avoids the use of hypoxia chambers or three gas incubators. These conditions of in vitro “Normoxia” have never been available in any other bioreactor on the market.

Petaka LOT Average DO = 1 mg/L (0.5 to 1.1 mg/L depending on cell type), equivalent to average 22.7 mmHg (Torr) in tissues or equivalent to average 3.11 %O2 in hypoxia chambers.
Petaka HOT Average DO = 3 mg/L (2.8 to 3.2 mg/L depending on cell type), equivalent to 66.5 mmHg in tissues or equivalent to 9.33 %O2 in hypoxia chambers

PetakaG3 2% Oxygen (Autonomous Physioxia chamber for cell culture)

Celartia introduces Petaka G3 the most physiologic hypoxia chamber in the world, mimicking as close as possible, how tissue cells in an organism are exposed to oxygen availability deficit.
Inside Celartia’s hypoxia chambers, cells are exposed to a progressive reduction of oxygen availability from 45 mmHg down to the lowest oxygen tensions recorded and published on living vertebrates: 13.32 mmHg.

Celartia’s hypoxia chambers Petaka G3 are completely autonomous.
Without hypoxia disturbances Celartia’s hypoxia chambers allow for bench top and open regular atmosphere use:
1. Continuous measurement of the oxygen reserve in the media.
2. Permanent evaluation of cellular oxygen consumption per cell and time.
3. Cell culture exposed to Hadron beams, electron beams and electromagnetic radiations maintaining the hypoxia level.
4. Celartia’s hypoxia chambers are closed and highly protected against contamination
5. Infinite media changes, sampling and special substances (and investigative drugs) additions to the media are performed in a regular laminar flow hood.
6. Laboratory bench Microscopy observations and time lapse video recording with regular microscopes, without glove boxes or any other ambient controlling devices
7. Can be incubated in any sort of temperature controlled device such as plain incubators, CO2 incubators, heat blocks or even water baths.
8. Celartia’s hypoxia chambers can be piled in arrays with efficient space and energy management
9. Celartia’s hypoxia chambers are disposable.

The scientific community agrees that mammal tissue hypoxia is a progressive onset status produced when the red blood cells can’t replace efficiently the oxygen of the interstitial media consumed by the tissue cells. This is exactly how Celartia’s hypoxia chambers regulate the hypoxia levels: in close interaction with the living cells. The proprietary micro-channel respiratory system incorporated in the device maintains under control the oxygen exchange, stabilizing the minimal level according to the type and number of cultured cells.

The discovery of the stress response to the oxygen deficit, specifically the cell response through the Hypoxia Induced Factor (HIF) and collaterals, opened a broad field of research in cell biology. As a consequence, new technologies were applied to produce environments where culturing cells with limited oxygen availability were possible.
The straight procedure used in industry has been the production of hermetic boxes (hypoxia chambers) filled and/or purged with the required gas mixture. Inside these boxes, traditional cell culture devices (flasks, bags and dishes) are installed and the whole assembly need to be introduced in incubators to achieve the required temperature.

This layer over layer arrangement implies many difficulties:
1. Restricted access to the cell culture device impairing:
a. Culture observation
b. Media changes
c. Adding supplements to the media

2. Gas composition Instability, due to:
a. The box needs to be opened for many required cell culture management operations.
b. Traumatic steps from normoxia to hypoxia in minutes
c. Gas decompression effect inside the cells
d. Uncontrolled gas diffusion through box materials and maintenance requirements

3. Only limited stepwise of hypoxia. There is no continuous transition from normoxia to hypoxia.
4. Exaggerated waste of space (volume)
5. Requirement of gas canisters as part of the assembly

Only a few highly sophisticated glove boxes available include a control unit program that permits gradual hypoxia in the chamber, where the cell culture devices are incubated; however, these complex devices are enormous, need great user training, are time consuming, need huge gas canisters, considerable power supply and laboratory space.

All of these restrictions and limitations are beat by Petaka G3 hypoxia chamber technology.

Outstandingly, Celartia’s Petaka G3 hypoxia chambers achieve this in the open atmosphere, inside a regular incubator, without accessory gas canisters, electronic sensors, feedback electronics or power supply.
Celartia’s hypoxia chamber’s manipulation is even easier than regular multi-well plates, and does not need special training to use them at the top level of efficiency.

Deep Hypoxia Experiments
Petaka offers the researcher complete freedom of sample manipulation even in very low oxygen hypoxic conditions, without breaking the hypoxic environment at any time.

Petaka, when packed in vacuum Mylar bags, it does not allow the passage of oxygen from the ambient atmosphere in the culture and the cells consume the media-dissolved oxygen in hours, creating a deep hypoxic condition. Cells can be grown and incubated in these conditions and a sample of the cell culture eventually withdrawn by punching the port directly through the Mylar bag, avoiding any exposure to higher oxygen concentrations.

To research cellular behavior under hypoxic or normoxic conditions, laboratories use controlled-environment incubators and glove boxes which offer the possibility of working in below ambient O2 concentration.

Glove boxes offer researchers the ability to incubate or perform sample manipulations without compromising the chamber’s gas-controlled environment. However Petaka does not need special incubators, glove boxes or any gas canisters.

No need for CO2 incubator 

Media based on Earle’s salts are buffered with a bicarbonate/carbonic acid system. These buffers rely upon the physiologically relevant pKa for carbonic acid and the equilibration of gaseous and media dissolved carbon dioxide to maintain the pH in a 5 to 10% CO2 incubator.

Petaka is designed to function without an additional CO2 environment. CO2 incubator is therefore not required, avoiding the need for CO2 cartridges. CO2 produced by the cell culture is also controlled. This is achieved by a gas transfer quenching system (GTQS) integrated into the device. As a result, the level of CO2 is maintained within the Petaka, allowing the pH to remain at a level compatible with the growth of 15 to 30 million cells.

Therefore, Petaka can be incubated in normal incubators without CO2 regulation, and cells grow in a closed and stable environment up to the limit of their own metabolism. These yields will vary according to each individual cell metabolism. We recommend that a culture test and a period of adaptation are implemented when a cell line is going to be cultured in Petaka.

Humidifiers Not Required.

Petaka is virtually vapor hermetic so does not require a humidity saturated incubator.

You may store pre-filled Petakas, ready to host cells, and have them prepared for immediate use. Refrigerated storage time is not limited by dehydration.
Elimination of saturated humidity in the incubator, water pan, and mist, reduces the risk of cross contamination in the incubator.

In Petaka, an integrated micro-channel system (MFS) protects the internal environment, avoiding unwanted water evaporation. Dehydration at 37ºC has been minimized, allowing culture incubation for 30 days at RH 10% with a final media dehydration below 10%.

Cryopreserve cells in a monolayer in the Petaka G3 - Superior to standard freezing in cryovials

The Petaka G3™ is the only cell culture device that also allows for convenient and direct cryopreservation inside the primary culture device. Cryo vials feature a very low surface-to-volume ratio. This means that the operator must use a slow freezing process while the glass transition progresses from the outside to the center of the vial. During this slow freezing period ice crystals may form inside of cells and damage them, leading to poor post-thaw viability. A preferred method is thin-film cryopreservation, only possible in the Petaka G3™. To freeze cells grown in a monolayer in the Petaka G3™ simply remove the culture medium, rinse cells briefly in freezing medium, then aspirate out excess freezing medium. This leaves only a thin film of freezing medium over a layer of cells no more than 20um thick. This thin film may then be immediately flash frozen (vitrified) for permanent cryopreservation. This process prevents trypsin exposure, and all risks and damage associated with the traditional slow-freezing process.

Cryovials are typically thawed in an unsanitary water bath for 1-2 minutes. This poses a risk to sterility and the extremely slow thawing technique leads to even more cell death. In contrast, thawing cells in a Petaka G3™ is achieved rapidly by filling it with warm medium. This means each cell thaws in just a fraction of a second as the warm medium covers the frozen cells; no water baths, running water, or sample transfers required. After adding warm medium, cells in the Petaka G3™ go directly to culture, with no reattachment or recovery time.

Up to 25 million attached cells may be frozen in a single Petaka G3™, using just 1ml of residual freezing medium. This means that when 24ml of warm medium is added to a Petaka G3™, the standard 10%DMSO is diluted to around 0.4%. This level of DMSO is no longer damaging to cells in culture, so
cells thawed in a Petaka G3™ do not need to be spun-down or washed during initial recovery and incubation. Cryopreservation in the Petaka G3™ means the whole process is faster, easier, carries less contamination risk, and leads to much higher post-thaw cell viability.

In vitro cell Dormancy or Pausing 

Using regular flasks, cells should be subcultured as soon as possible when they reach a confluent monolayer state or saturate the carriers.

With Petaka, grown cell cultures can be maintained alive at room temperature, without dehydration risk, for long time periods.This facilitates cell culture management and maintenance routines; and provides additional advantages for cell biology and toxicology research (see “Petaka Quick Protocols” for conditions and methods, such as protocols 4 and 8 – “Keeping Cells in In-Vitro Dormancy”).

Cell conservation in dormant state is favored by transferring cultures from 37° C to 22° C when cells are in a slow progressing cell cycle. This is achieved in an 80% confluent state.
Under low temperature and low pH, cells reduce DNA replication practically to zero, and remain in a suspended animation-like state (a certain accumulation of cells in G2 phase could be expected).The duration of the dormancy period is dependent on the cell type. At a minimum, 7 days survival is common for most cells, while others could live for more than 200 days.

Specifications

Dimensions:
Inches: 5.03 x 3.38 x 0.21
mm: 127.8 x 85.7 x 5.45
Maximum media volume: 25 mL 
Maximum cell culture surface: 150 cm2

Characterization:
The cell culture device PetakaG3 is an innovative bioreactor designed for mammalian cell culture in physiologic tissue conditions.
The distinctive feature of PetakaG3 is that being a disposable device incorporates an automatic gas diffusion control system (AGD-CS) that works interactively with the cells growing and living inside, WITHOUT HYPOXIA CHAMBERS, GLOVE BOXES OR THREE-GAS INCUBATORS.
For cells cultured in PetakaG3 and incubated in regular 21% oxygen atmosphere the AGD-CS provides: 

  •  Media dissolved oxygen concentration within the limits of the physiologic oxygen tensions in physiologic tissues (from 75 mmHg down to 15 mmHg) monitorable with a dissolved oxygen fluorescent sensor dot. Because this advanced trait PetakaG3 is the safest, smallest, unique portable disposable hypoxia chamber, also available with integrated DO sensor.
  • Media dissolved CO2 concentration, enough to maintain a physiologic pH without external extra-supply of carbon dioxide gas.  
  • Controlled media evaporation even at 2% to 10% atmospheric Relative Humidity (RH). PetakaG3 is available in several options for culturing all kinds of cells from vertebrate and invertebrate organisms, especially suited for stem cells, primary cultures and tumor cells. PetakaG3 LOT & PetakaG3 HOT with hydrophilic surface developed for anchored dependent cells growth and PetakaG3 FLAT with a hydrophobic surface, not permissive of cell adhesion, for non adherent cells growth and 3D cellular clusters and aggregates. PetakaG3-Matrix is available carrying different surface molecular matrices, such as Collagen, Vitronectin, Matrigel, etc. and also structural matrices (nano-fibers mesh), or combinations of structural and molecular. 
  • PetakaG3 allows cell cultures of two different cell types in the same device. Both cell layers are separated by 3mm of media, which is appropriate to study intercellular interactions through diffusible factors and exosome exchange. PetakaG3 is ideal for transfection of both cell layers with two different gene constructs in the same device with negligible cross transfection

Frequently Asked Questions