Method of cell cultures and device for implementing it
The invention generally provides a cell culture vessel having at least one first zone and at least one second zone, wherein the first zone is a transfer zone for a culture medium which essentially contains no cells and the second zone is a cell culture zone. The invention further includes methods utilizing the cell culture vessel.
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 for transferring the culture medium essentially containing no cells and said second zone is for culturing cells, means of circulating the culture medium, allowing circulation of the culture medium through the second zone, said second 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, and further comprising at least one third and at least one fourth zone, wherein both are essentially free from cells, wherein said third zone is in medium communication with the first and second zones and wherein said fourth zone is in medium communication with the second zone and with the first zone via the means for circulating the culture medium, and in that the means for circulating is adapted to circulate the culture medium from bottom to top in said second zone, wherein said third zone includes an overflow so that the culture medium overflows from the first zone to the third zone.
2. The device according to claim 1 , 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.
3. The cell culture device according to claim 1 , wherein the third zone is a zone located entirely below the second zone and entirely above the first zone.
4. The cell culture device according to claim 2 or 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.
5. The cell culture device according to claim 2 , 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 means for circulating.
6. The cell culture device according to claim 5 , wherein the means for circulating is 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.
7. The cell culture device according to claim 5 , further comprising a head module (m t ), said head module (m t ) comprising at least the cover.
8. 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.
9. The cell culture device according to claim 1 , wherein said essentially vertical or inclined flow wall comprises a hydrophilic membrane.
10. The cell culture device according to claim 1 , further comprising at least one gas inlet orifice and one gas outlet orifice.
11. 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 zone.
12. The cell culture device according to claim 5 , wherein the means for circulating is 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 connector and in its bottom part a second connector and wherein said base module (m 0 ) also comprises in a top part (m 0b ) a first complementary connector and said head module (m t ) also comprises in its bottom part a second complementary connector for producing a stacking sequence from bottom to top of a base module (m.sub. 0 ), at least one culture module (m 0 . . . m n ) and a head module (m t ).
13. The device according to claim 12 , wherein said connectors are adapted for forming said stacking sequence in a gas- and liquid-tight manner.