The Passage

In cellular culture, the passage is the process of subculture of animal cells. It is in general made to produce a great number of cells starting from existing pre cells.

In laboratory, adhering cells of mammal are cultivated in of Flasks or of limp of varied $petri of diameter, with SVF and medium DMEM and incubated with 37°C in an atmosphere of carbon dioxide 5% and in the presence of water to have a constant moisture. The cells in culture then will grow until entirely papering the bottom of the flask or of limps of Petri. In normal weather, for example for a culture of cells of the type HeLa recovering 10% of the surface of culture, it will be necessary between two and three days to reach 100% of Confluence. If nothing is considered at this stage, food starts to rarefy and the cells start to die; more cells which are brought to develop without adhering itself will die more quickly and could not be mended. A passage is thus nécessaire' i.e. that one will mend the cells in a news flask with culture medium nine.

Handling proceeds thus. The impoverished medium is removed, generally using a pipette connected to a liquid jet vacuum pump. The cells are washed with a Tampon phosphates saline called PBS (saline phosphate buffered), then a solution of Trypsine is added in order to take off the cells of the bottom of the flask (to let act 5 minutes with 37°C in the incubator). Then one adds a solution of plug phosphates saline and one operates aspirations repressions with a pipette for to the maximum individualizing the cells. One continues by taking a tenth of the cellular volume of suspension contained in the flask, and one deposits them in a sterile news flask new medium DMEM. One finishes while placing the flask in the incubator. And the cycle starts again until the next passage. The remainder of cells is eliminated in a solution chlorinated to kill these last.

It should be noted that for better results, the passage will have to be carried out before arriving at 100% of junction.

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