Carrier for undifferentiated cell culture and subculture method thereof
View Patent ↗A colony (cell mass) proliferated under the undifferentiated state is obtained by using a carrier for cell culture in which two or more of a concavity having a porous body in a surface are arranged on a substrate surface in the form of a matrix, inoculating an undifferentiated cell on at least one concavity of the carrier for culture and carrying out culture.
1. A carrier for cell culture, comprising concavities arranged in line direction and column direction on a surface of the carrier, wherein the whole carrier, including the surface, consists of a porous body and wherein a cultured cell is capable of adhering to the inside of each of the concavities, and wherein the porous body has a pore size of from 10 nm to 2,000 nm, and wherein the porous body consists of at least one material selected from the group consisting of zirconia, yttria, titania, alumina, hydroxyapatite, and β-calcium tertiary phosphate, and
wherein the concavities each have a diameter from 10 μm to 1,000 μm and a depth of from 30 μm to 1,000 μm.
2. The carrier according to claim 1 , wherein the concavities have a bottom of a hemispheric shape.
3. The carrier according to claim 1 , wherein the concavities each have a diameter of from 100 μm to 400 μm and a depth of from 50 μm to 400 μm, wherein the concavities are each capable of culturing a mouse ES cell.
4. The carrier according to claim 1 , wherein the concavities each have a diameter of from 250 μm to 1,000 μm and a depth of from 125 μm to 1,000 μm, wherein the concavities are each capable of culturing a human ES cell.
5. The carrier according to claim 1 , wherein the porous body is selected from the group consisting of zirconia and alumina.
6. A method for culturing a cell, which comprises using the carrier of claim 1 , inoculating an undifferentiated cell on at least one concavity of the carrier for cell culture and carrying out a culture.
7. A method for culturing a cell, which comprises carrying out a subculture by repeating the method for culturing described in claim 6 , by separating a cell mass obtained by culture into a single cell or a small cell population, and using the cell obtained by exfoliation from the carrier for culture.
8. The method for culturing a cell according to claim 7 , wherein separating the cell mass is carried out by enzyme treatment and pipetting.
9. A method for culturing a cell, which comprises carrying out suspension culture by exfoliating the cell mass obtained in claim 7 from the carrier for culture in the state of cell mass, and using the cell mass.
10. The method for culturing a cell according to claim 9 , wherein exfoliating the cell mass from the carrier for cell culture is carried out by pipetting alone.