In vitro culture medium for cells, tissues, and organs of amphibians

Currently, in research and experiments, during which amphibian cell and tissue cultures are carried out, appropriately diluted media mixtures developed for mammals are used. Unfortunately, some components of such media, despite adequate dilution, may have a toxic effect on amphibian cells. On the other hand, after diluting the media originally intended for mammalian cell and tissue culture, in order to adapt them to amphibian cell culture, the concentrations of all substances are reduced, which means that some of them stop working. This clearly indicates that the dilution of the medium developed for the culture of mammalian cells is not sufficient to ensure good conditions for culturing the cells, tissues, and organs of amphibians. Therefore, the development of a medium, the composition of which in terms of active substances and their concentrations would be formulated for the culture of amphibian cells, is desirable.

The subject of the offer is a specialized medium for the in vitro culture of cells, tissues, and organs of amphibians. Thanks to the unique composition of active substances and refined their concentrations to the requirements of amphibian cells, the use of medium enables long-term culture as well as a high level of cell proliferation and reduction of cell mortality.

The final stage of the research on invention was the comparison of the amphibian cell and organs culture carried out using the developed medium in relation to the cultures carried out with most commonly used mixtures of mediums developed for the cultivation of mammalian cells.

The following media were used in comparative studies:

A - (67% L15 + 10% FBS) (Sinzelle et al. 2012);

B - (33,3 mL L15, 33,3 mL RPMI1640+HEPES + 10%FBS + 1,33 mg/Ml sodium bicarbonate + 1 mM sodium pyruvate) (Tlapakova et al. 2016);

C- (66% L15 + 10%FBS) (according to Stukenberg Lab);

D - (33,3 mL L15, 33,3 mL RPMI1640+HEPES + 6,65%FBS + 26 mL water) (Piprek et al. 2013);

T- Developed medium

Comparative research has shown that:

- the developed medium enables the cultivation of amphibian cells in vitro for the longest period of time (up to 30 days) (Figure 1)

Figure1

- culture of cells enzymatically isolated from the Xenopus laevis amphibian, carried out with the developed medium, showed the highest number of proliferating cells and the lowest number of necrotic cells (Figure 2)

Figure 2

- cells isolated from gonads of Xenopus laevis tadpoles cultured using the developed medium showed the highest level of cell proliferation and the lowest level of necrosis (Figure 3)

Figure 3

Intellectual property status

Patent already applied for

Patent application number : P 428393

Where : Poland

Current development status

Laboratory prototypes

Desired business relationship

Technology selling

Patent licensing

Technology development

Adaptation of technology to other markets