Cell culture apparatus, apparatus for long-term observation of cell culture, method for long-term cell culture, and method for long-term observation of cell culture

摘要

Provided is a cell culture apparatus, an apparatus for long-term observation of cell culture, a method for long-term cell culture, and a method for long-term observation of cell culture, which are capable of continuously culturing and observing cells under uniform environmental conditions without aging of the cultured cells' physiological state, and also tracing the history of a cell. The cell culture apparatus comprises a cell culture substrate, a semipermeable membrane, and a culture solution supplier. The cell culture substrate has on a surface a narrow culture groove for holding and culturing cells and wide flow grooves for discarding the cells held and cultured in the culture groove. Both ends of the culture groove are connected to the flow grooves being larger in width and depth than the culture groove. The semipermeable membrane covers the culture groove and the flow grooves. Finally, the culture solution supplier continuously supplies culture solution.

主权项

1. A cell culture apparatus for culturing cells, comprising a cell culture substrate, a semipermeable membrane, a liquid feeding pad and a culture solution supply means, wherein
the cell culture substrate comprises:
(1) a culture groove capable of holding and culturing the cells, wherein the culture groove has a depth 1 to 2 times the size of a desired cell and a width 1 to 3 times the size of the desired cell,(2) a first flow groove having a width 10 to 100 times the width of the culture groove and a depth 1.5 to 50 times the depth of the culture groove, wherein a side of the first flow groove is attached to a first end of the culture groove at an angle of the flow groove relative to the culture groove of 90°±15°, and(3) a second flow groove having a width 10 to 100 times the width of the culture groove and a depth 1.5 to 50 times the depth of the culture groove, wherein a side of the second flow groove is attached to a second end of the culture groove at an angle of the flow groove relative to the culture groove of 90°±15°; the semipermeable membrane covers the culture groove and the flow grooves of the cell culture substrate; the liquid feeding pad is disposed on the substrate and forms a space over the culture groove, the first and second flow grooves and the semipermeable membrane, and the liquid feeding pad is adhered to a periphery of an area of the substrate comprising the culture groove and the first and second flow grooves; the culture solution supply means includes a liquid feeding pump and a waste liquid tank; and the cell culture apparatus is configured to continuously culture cells for 200 generations or more by continuously supplying culture solution through the first and second flow grooves to the culture groove and continuously removing culture solution and a portion of cells from the first and second ends of the culture groove through the first and second flow grooves to the waste liquid tank.