Cell culture kit, screening method, and method of manufacturing cell culture kit
To provide a cell culture kit including cultured living cells of various donors, and a manufacturing method thereof. The cell culture kit includes a culture plate and living cells cultured thereon. The culture plate includes a plurality of microchambers ( 33 ) and living cells derived from various donors are adhered to surfaces of the plurality of microchambers ( 33 ). Specifically, living cells D 1 , D 2 , and D 3 derived from various donors are adhered to the plurality of microchambers ( 33 ). In each microchamber ( 33 ), living cells derived from one donor or living cells derived from various donors may be cultured. The living cells derived from various donors are adhered and cultured in the cell culture kit as a whole, which makes it possible to provide a cell culture kit to conduct a test using cells derived from various donors.
1. A method of manufacturing a cell culture kit, comprising:
obtaining liver cells from at least two different human donors;
seeding the liver cells in a plurality of micro spaces contained in each of a plurality of chambers in a cell culture plate, with liver cells from only one of the different human donors seeded in at least one of the micro spaces, and liver cells from at least two of the different human donors seeded in at least one of the micro spaces separate from a micro space containing liver cells from only one of the different human donors, and the at least one micro space containing cells from only one of the different human donors is adjacent to the micro space that contains cells from the at least two of the different human donors;
adhering the seeded cells to the plurality of micro spaces; and
culturing the seeded liver cells within the micro spaces in a culture medium,
wherein the plurality of micro spaces each have a bottom area of 0.01 mm 2 to 0.1 mm 2 , a depth of 25 μm to 150 μm, and walls partitioning the micro spaces with each side wall having a width in the range of 3 μm to 15 μm,
wherein an inorganic hydrophilic membrane is uniformly formed on the surface of the chambers, and
wherein an organic film made of extracellular matrix suitable for cultured cells is disposed on the inorganic hydrophilic membrane, and the seeded liver cells adhere to the film.
2. The method of claim 1 , wherein liver cells from at least two of the different human donors are seeded in at least one micro space adjacent to the micro space that comprises the liver cells from the at least two of the different human donors.
3. The method of claim 1 , wherein the cells comprise a liver stem cell.
4. The method of claim 1 , wherein the cells comprise at least one selected from the group consisting of a cell differentiated from an embryonic stem (ES) cell and a cell differentiated from an induced pluripotent stem (iPS)-cell.
5. The method of claim 1 , wherein the cells aggregate and form a three-dimensional structure during culturing.
6. The method of claim 1 , wherein the cells are seeded in the plurality of micro spaces at a density of 1×10 2 to 1×10 6 cells/cm 2 .
7. The method of claim 1 , wherein the cells form a cell aggregate in each of the plurality of micro spaces.
8. The method of claim 7 , wherein the cell aggregate has a diameter of 30 to 200 μm.
9. The method of claim 1 , wherein the cells comprise a liver precursor cell.
10. The method of claim 1 , wherein the liver cells are isolated from hepatic tissues of the at least two different human donors.
11. The method of claim 1 , wherein the culture medium covers the plurality of micro spaces.
12. The method of claim 1 , further comprising before seeding the cells, obtaining the cells from more than two of the different human donors.
13. The method of claim 1 , wherein each chamber further comprises a plurality of spots, the plurality of spots are regions containing the plurality of micro spaces, and the different spots containing the micro spaces are partitioned from each other by a side wall.