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Normoxia conditions in Petaka... tissue-like conditions!

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There are many sophisticated (and with that, we usually presume expensive) instruments on the market enabling researchers to achieve the essential physiological conditions necessary to truly study cell behavior in vitro as in vivo.
But even these instruments use untried & untested physiological conditions, replacing oxygen with nitrogen or helium in an attempt to replicate the mammalian or human hosts.

Petaka, a simple, yet highly innovative plastic labware device, enables the cells themselves to auto-regulate the levels of oxygen & CO2 in the media without using any external instrumentation or apparatus.

 

Oxygen concentrations... Normoxia with Petaka

Achieving the correct concentration design is no longer a problem
 
Petaka provides the cells with a gradually decreasing oxygen concentration in the media, within the physiological limits of living tissues (Graph 1). Cell culture may begin with a concentration equivalent to that of arterial blood (OP 75 mmHg), high enough to promote exponential cell growth. The concentration is then progressively reduced to that of embryonic development (OP 15 mmHg), avoiding cell damage and facilitating cell differentiation.
 

No need for CO2 incubator to cultivate cells in Petaka

CO2 cartridges are no longer necessary
 
Petaka is designed to allow very low amounts of CO2 in and out . Additional COis therefore not required, avoiding the need for CO2 cartridges. COproduced by the cell culture is also controlled. This is achieved by a gas transfer quenching system (GTQS) integrated in the device. As a result the level of COis maintained within the Petaka, allowing the pH to remain at a level compatible with the growth of 16 to 22 million cells. Therefore, Petaka can be incubated in normal incubators without CO2, 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 grown in Petaka.


Hypoxia experiments without hypoxia chamber

Without special incubators

Low oxygen concentration in the cell culture media has a significant impact on cell physiology and differentiation. Cells kept in traditional hyperoxic culture conditions provided by media in contact with ambient air containing 20 % oxygen behave very differently. 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. Controlled-environment incubators only allow the incubation of tissue culture flasks in a controlled oxygen atmospheres. Glove boxes offer researchers the ability to incubate or perform sample manipulations without compromising the chamber’s gas-controlled environment. However the manipulation of cell cultures in glove boxes can be difficult and uncomfortable.
Petaka does not need special incubators or glove boxes. Petaka offers the researcher complete freedom of sample manipulation even in very low oxygen hypoxic conditions, without breaking the hypoxic environment at any time.

No lost of humidity in Petaka!

No more water pan or humidifier inside the incubator

Petaka is virtually vapor hermetic
Petaka does not require a humidity saturated incubator. You may store pre-filled Petakas, ready to host cells, and have them available for immediate use.

Increased protection against incubator cross contamination
Elimination of saturated humidity in the incubator, water pan, and mist, reduces the risk of cross contamination in the incubator.

Refrigerated storage time is not limited by dehydration
Petaka pre-filled with media can be stored for a long time in most refrigerators at 4ºC without the risk of dehydration. The refrigerator type should be considered before storage to avoid unexpected dehydration rates.

Contamination protected cell culture device

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 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. All these features, and the fact that Petaka does not require a humid incubation environment, make it one of the most contamination protected cell culture devices available.

Cell transport of living cells at normal temperature

With the Petaka you can transport live cells for long distances at normal temperatures. Petaka does not need dry ice or refrigeration to transport cells. Its slim shape permits easy packaging and protection. Padded envelopes are ideal for shipping one or several 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.

 

In Vitro cell dormancy in Petaka

One main objective of Petaka technology is to provide freedom to researchers by reducing demanding time consuming routines. 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.

 

 

Protocols

Physiological relevance is an essential feature for any laboratory looking to work with cells replicating in vivo such as stem cell research for regenerative applications, work with primary cultures & cancer research & other potentially deadly diseases etc.  The unique shape & design of the Petaka allows researchers to conduct a series of applications or protocols on their cell culture without ever having to remove those cells from the device at all.

Culturing, washing, magnetic sorting, magneto-transfection, pelleting or separating, enzyme-free harvesting, shipping & freezing can all be performed within the same device without having to transfer cells to any other tube, vial or instrument. This as a consequence almost totally eliminates the likelihood of contamination & saves a lot of time in avoiding unnecessary protocols.

So in nutshell, this is the philosophy of Petaka: to strive to develop more efficient protocols, to save time & reduce consumption & ultimately facilitate better tools & conditions for researchers.

 

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