METHOD OF CELL CULTURES AND DEVICE FOR IMPLEMENTING IT

1 . A cell culture device comprising: a culture vessel provided with a cover, wherein there is situated at least one first zone and at least one second zone, wherein said first zone is a transfer zone for the culture medium essentially containing no cells and said second zone is a cell culture zone, means of circulating the culture medium, allowing circulation of the culture medium through the culture zone, said culture zone comprising a bottom wall and a top wall, wherein each wall is provided with orifices allowing a transfer of culture medium essentially free from cells, characterised in that: it also comprises at least one third and at least one fourth zone, wherein both are culture medium transfer zones essentially free from cells, wherein said third zone is in medium communication with the first and second zone and wherein said fourth zone is in medium communication with the second zone and with the first zone via the culture medium circulation means, and in that the culture medium circulation means allow a circulation of the culture medium from bottom to top in said second culture zone.

2 . The device according to claim 1 , wherein said third zone, has an overflow so that the culture medium overflows from the first culture medium transfer zone to the third culture medium transfer zone.

3 . The device according to claim 2 , wherein said third zone is a zone internal to said second zone and external to said first zone and said fourth zone is a zone external to said second zone.

4 . The cell culture device according to claim 2 , wherein the third zone is a zone located entirely below the second zone and entirely above the first zone.

5 . The cell culture device according to claim 3 , further comprising flow redistributing elements in the third zone or closed regions in the bottom wall of the second zone for providing a homogenous flow of medium into said second zone.

6 . The cell culture device according to claim 3 , comprising a series of culture modules (m 1 , . . . , m n ), each module comprising said first zone, said second zone, said third zone and said fourth zone, and wherein the adjacent modules in said series of culture modules are in medium communication, and said first zone and said fourth zone of each module are in communication with said circulation means, directly or indirectly.

7 . The cell culture device according to claim 6 , wherein the circulation means are confined in a base module (m 0 ), and wherein said base module (m 0 ) is in medium communication with at least one first medium transfer zone and at least one fourth medium transfer zone, directly or indirectly.

8 . The cell culture device according to claim 6 , further comprising a head module (m t ), said head module (m t ) comprising at least the cover.

9 . The cell culture device according to claim 1 , wherein said at least one fourth zone comprises at least one essentially vertical or inclined flow wall.

10 . The cell culture device according to claim 1 , wherein said essentially vertical or inclined flow wall comprises a hydrophilic membrane.

11 . The cell culture device according to claim 1 , further comprising at least one gas inlet orifice and one gas outlet orifice.

12 . The cell culture device according to claim 1 , wherein the cover of the culture vessel is connected to at least part of said top wall of at least one second culture zone.

13 . The cell culture device according to claim 6 , wherein the circulation means are confined in a base module (m 0 ), wherein said base module (m 0 ) is in medium communication with at least one first medium transfer zone and at least one fourth medium transfer zone, directly or indirectly, wherein each culture module (m 0 , . . . , m n ) comprises in its top part a first fixing means and in its bottom part a second fixing means and wherein said base module (m 0 ) also comprises in a top part (m 0b ) a first fixing means and said head module (m t ) also comprises in its bottom part a second fixing means, wherein said first fixing means and said second fixing means are complementary fixing means for producing a stacking sequence from bottom to top of a base module (m 0 ), at least one culture module (m 0 , . . . , m n ) and a head module (m t ).

14 . The device according to claim 13 , wherein said first and said second fixing means comprise means for producing said stacking sequence in a gas- and liquid-tight manner.

15 . A method of culturing cells in a culture vessel with culture medium circulation, comprising the steps of: introduction of culture medium (M) into culture medium circulation means, discharge of culture medium (M) from said culture medium circulation means, at least one first transfer of culture medium (M) into a first culture medium upward transfer zone, at least one second transfer of culture medium (M) into a second cell culture zone, characterised in that it also comprises: at least one third culture medium transfer subsequent to said first culture medium transfer into a third culture medium transfer zone, by overflowing from the first culture medium transfer zone to the third culture medium transfer zone, at least one fourth culture medium transfer subsequent to said second culture medium transfer into a fourth culture medium transfer zone, by overflowing from the second culture zone into the fourth culture medium transfer zone, and in that said second culture medium transfer is a downward culture medium transfer.

16 . The method of culturing cells according to claim 15 , also comprising oxygenation of the culture medium during one or more of said transfers.

17 . The method of culturing cells according to claim 15 , wherein the oxygenation is carried out during the fourth culture medium transfer, and wherein said fourth culture medium transfer is a flow of said culture medium (M) along a flow wall.

18 . The method of culturing cells according to claim 15 , wherein said flow of the culture medium (M) is a flow along a hydrophilic wall.