Although occasionally taken for granted, a tightly controlled, reliable incubation system can be one of the most valuable pieces of equipment in a laboratory. The ability to culture a healthy cell line can make the difference between a successful and failed investigation, and influence the results of research projects. One prime example of this is the IVF research taking place at the Karolinska Institute in Sweden, where PHCbi incubators are being used for cell cultivation.
The Karolinska Institute
With assisted reproduction now increasingly widespread and scientific technology constantly evolving, it is no surprise that the improvement of IVF is a topic for continuing investigation. It has been estimated that today there has been more than 5 million IVF births worldwide.
The continuing development of easier methods has increased interest and introduced opportunities to make IVF more affordable, successful and available to the wider population. Some of this research is being undertaken at the Karolinska Institute, the biggest medical university in Sweden.
The prestigious Institute has a large stem cell programme containing many groups, one of which is led by Professor Outi Hovatta and dedicated to making IVF available to women for whom it may not have been an option. To achieve this, the team is investigating the maturation of human ovarian follicles and oocytes from cryostored human ovarian tissue. When there are risks to implanting tissue directly back into the ovary of a woman who has lost her eggs, perhaps due to chemotherapy or genetic causes, this could provide a viable alternative.
To support the effective development of this innovative and ground-breaking technique, the Karolinska Institute has opted for a high quality, stringently controlled incubation system. Culturing oocytes from primordial follicles to full maturity is a long and delicate process that can be severely affected by the cell’s environment. Therefore the institute depends heavily on its incubation systems.
PHCbi’s innovative incubator technologies are ensuring that the required levels of temperature and environmental control are achieved.
Replicating an in vivo environment
The Karolinska Institute uses around twenty IncuSafe MCO-5M multi gas incubators in its laboratories. These small personal appliances are well suited for applications involving professional cell culturing, due to their precise CO2 and O2 level controls, reliable decontamination and temperature stability, all of which assist in creating a stable environment for cells. This level of environmental control is vitally important in IVF research, and was a major consideration for the Karolinska Institute when specifying their systems.
In order to culture mature oocytes, very precise conditions are required. For example, the temperature must remain at a constant 37ºC and reduced oxygen concentrations of 5% must be maintained, reflecting the natural in vivo conditions of the cells. This is essential to guarantee the viability of the mature cells and maximise their success in future IVF procedures.
Innovative technologies for an innovative application
The Karolinska Institute’s cell cultures are always kept these optimum conditions, whether they are situated by the incubator door or right at the back of the chamber. This is initially guaranteed by Direct Heat and Air Jacket, which ensures and maintains the required temperature of 37ºC. In addition, gentle fan circulation reduces any temperature gradient throughout the chamber.
In order to provide physiological conditions for the cells in culture, the incubators also achieve stable gas concentrations through a combination of exclusive technologies. A long-life zirconia oxygen sensor upholds sub-ambient oxygen levels from 1% to 18%.
An automatic N2 cylinder switchover system is also included, which switches from a primary to a secondary cylinder when recovery times exceed a given limit. In this way there are no large variations in gas concentration, and so no risk to the biological processes of the maturing ovarian follicles.
A centralised microprocessor controller complete with alarm, programming, calibration and diagnostic protocols supervises all operations within the incubator, so that internal conditions in the incubators are constantly monitored and controlled. Should there ever be an issue, researchers are alerted immediately. In this way, the oocytes receive with ultimate levels of security.
The inclusion of multiple decontamination systems further guarantees in vitro performance by including inCu saFe® and SafeCell® UV integrated contamination control technology. This powerful combination protects against both airborne and waterborne contaminants, and prevents the growth of moulds, fungi and bacteria with constant germicidal protection. In this innovative application, where cells are typically one-of-a-kind, this provides additional peace of mind to the team at the Karolinska Institute.
With all these technologies implemented in a single piece of equipment, the Karolinska Institute’s PHCbi incubation system is ideal for the pioneering research taking place. By providing a stable environment, primordial follicles can be cultured successfully into mature oocytes for IVF. Although the cells have yet to reach the ideal size for implantation (120 μm), ethics approval has already been given to move this research into the next stage. “The cell environments created by the PHCbi incubators are very stable and well controlled,“ commented Professor Hovatta. “In our research, the cell lines are completely unique, so we cannot take any risks during their cultivation. By developing this technique, we are confident that we can demonstrate that the resulting oocytes and embryos are normal and healthy, and eventually progress to clinical applications.” Professor Hovatta’s final aim is to have this technique introduced into clinical settings, making IVF and assisted reproduction a viable option for even more women looking to become pregnant.