Methods of producing three-dimensional cellular tissues
AssigneeToppan Holdings Inc.
InventorsIsana KONDO, Rii MORIMURA, Shiro KITANO
Life of this patent
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Abstract
A method of producing three-dimensional cellular tissues includes mixing stromal cells with a cationic substance, an extracellular matrix component and a polyelectrolyte to obtain a mixture; removing a liquid portion from the mixture to obtain a cell aggregate; culturing the cell aggregate in a medium to obtain a three-dimensional cellular tissue; and culturing the three-dimensional cellular tissue in a medium containing ascorbic acid and Transforming Growth Factor-β (TGF-β), wherein a thickness of the three-dimensional cellular tissue remains greater than 50 μm for at least 3 days after culturing the three-dimensional cellular tissue in the medium containing ascorbic acid and TGF-β.
Claims11
A method of producing three-dimensional cellular tissues, comprising:
- mixing stromal cells with a cationic substance, an extracellular matrix component and a polyelectrolyte;
- removing a liquid portion from a mixture of the stromal cells, the cationic substance, the extracellular matrix component and the polyelectrolyte such that a cell aggregate is obtained;
- culturing the cell aggregate in a first medium such that a three-dimensional cellular tissue is obtained;
- and
- culturing the three-dimensional cellular tissue in a second medium comprising ascorbic acid and Transforming Growth Factor-β,
- wherein a thickness of the three-dimensional cellular tissue remains greater than 50 μm for at least 3 days after culturing the three-dimensional cellular tissue in the second medium comprising ascorbic acid and Transforming Growth Factor-β.
A method of producing three-dimensional cellular tissues, comprising:
- mixing stromal cells with a cationic substance, an extracellular matrix component and a polyelectrolyte such that a first mixture is obtained;
- removing a liquid portion from the first mixture of the stromal cells, the cationic substance, the extracellular matrix component and the polyelectrolyte such that a first cell aggregate is obtained;
- culturing the first cell aggregate in a first medium such that a first three-dimensional cellular tissue is obtained;
- disposing target cells on the first three-dimensional cellular tissue;
- mixing stromal cells with a cationic substance, an extracellular matrix component and a polyelectrolyte such that a second mixture is obtained;
- removing a liquid portion from the second mixture of the stromal cells, the cationic substance, the extracellular matrix component and the polyelectrolyte such that a second cell aggregate is obtained;
- disposing the second cell aggregate such that the second cell aggregate is in contact with target cells;
- and
- culturing the second cell aggregate in a second medium comprising ascorbic acid and Transforming Growth Factor-β such that a second three-dimensional cellular tissue is obtained,
- wherein a total thickness of the first three-dimensional cellular tissue, the target cells and the second three-dimensional cellular tissue remains greater than 50 μm for at least 3 days after culturing the second three-dimensional cellular tissue in the medium containing ascorbic acid and Transforming Growth Factor-β.
Dependencies are inferred from the claim text.