Patent ID: 9587216
Date: 2017-03-07
CPC Classifications: C12M

Claim:
1. A cell culture method characterized in that it is implemented in a system comprising: at least one device suitable for culturing cells, the device comprising: said method comprising: a) filling the pre-chamber with a culture medium under initial physicochemical conditions with a predetermined volume of liquid, b) introducing cells in the pre-chamber through the cell inlet, c) passing the culture medium with the cells to the at least one bioreactor chamber, d) leaving the cells to settle and leaving them to stand for a predetermined time until they adhere to the at least one inner surface suitable for cell culture the physicochemical properties of which cause it to behave in such a way that it favors cell adhesion under one or several initial physicochemical conditions, e) circulating culture medium between said at least one bioreactor chamber and said at least one pre-chamber through the fluid inlet-outlet means, f) once a target number of cells is obtained or a critical density is reached in the at least one bioreactor chamber used for the culture, changing at least one of the initial physicochemical conditions of the culture medium to modified physicochemical conditions under which cell adhesion is prevented, such that the volume remaining within the device under modified physicochemical conditions is located in the bioreactor chambers which have been used for cell amplification, g) passing the culture medium with the cells to the pre-chamber, and h) either extracting the culture medium with the cells in suspension through outlet means, if the target number of cells has been reached; or  introducing additional culture medium in the pre-chamber and repeating the steps starting from step c), if the target number of cells has not been reached.