Method for producing cell aggregate
View Patent ↗Disclosed is a method for producing a cell aggregate having bile canaliculi, including a contact step of bringing cells including hepatocytes into contact with heparin in an aqueous medium and a culture step of culturing the cells brought in contact with the heparin to form the cell aggregate. A concentration of heparin in the aqueous medium when it is brought into contact with the cells may be 0.5 mg/mL or more.
1 . A method for producing a cell aggregate comprising bile canaliculi, comprising:
a contact step of bringing cells comprising hepatocytes into contact with heparin in an aqueous medium; and
a culture step of culturing the cells brought in contact with the heparin to form the cell aggregate,
wherein a concentration of heparin in the aqueous medium when brought into contact with the cells is 0.5 mg/mL or more.
2 . The method according to claim 1 ,
wherein the hepatocytes are mature hepatocytes.
3 . The method according to claim 1 ,
wherein the proportion of the number of hepatocytes in the total number of cells is 65% or more.
4 . The method according to claim 1 ,
wherein, in the contact step, the cells are additionally brought into contact with extracellular matrix components.
5 . The method according to claim 4 ,
wherein the extracellular matrix components are collagen components.
6 . The method according to claim 1 ,
wherein the expression intensity percentage of multidrug resistance-associated protein 2 (MRP2) in the cell aggregate represented by the following Formula (1) is 120% or more,
expression intensity percentage of MRP2=X/Y×100 (1)
wherein in Formula (1),
X represents an expression intensity of MRP2 in the cell aggregate formed in the culture step, and
Y represents an expression intensity of a control cell aggregate formed by culturing cells comprising hepatocytes without contact with heparin and extracellular matrix components.
7 . The method according to claim 1 ,
wherein the concentration of heparin in the aqueous medium when brought into contact with the cells is 0.5 mg/mL or more and 12.0 mg/mL or less.
8 . The method according to claim 1 ,
wherein the concentration of heparin in the aqueous medium when brought into contact with the cells is 0.5 mg/mL or more and 5.0 mg/mL or less.