Method of producing frozen renal cells and frozen renal cell
AssigneeNikkiso Co., Ltd.
InventorEtsushi Takahashi
Life of this patent
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Abstract
A method of producing proximal tubule epithelial cells includes recovering the cells as first aggregates, freezing the aggregates while substantially maintaining their aggregated state, thawing the frozen cells, and culturing the thawed cells. The thawed cells are cultured in a non-adherent state on a culture vessel subjected to a cell non-adhesion treatment for 7 days or more, thereby forming second aggregates. When cultured for 7 days or more after thawing, the cells exhibit gene expression levels of at least 10% of those in human renal cortex cells for at least one gene selected from AQP1, CD13, SGLT2, Na/K ATPase, URAT1, PEPT1, MDR1, OAT1, OCT2, OCTN2, E-cadherin, and ZO-1.
Claims10
1-11. (canceled)
A method of producing proximal tubule epithelial cells comprising:
- recovering proximal tubule epithelial cells as first aggregates;
- freezing the recovered proximal tubule epithelial cells;
- thawing the frozen proximal tubule epithelial cells;
- and
- culturing the thawed proximal tubule epithelial cells;
- wherein the first aggregates are frozen while an aggregated state of the first aggregates is substantially maintained in the freezing step;
- wherein the cultured thawed proximal tubule epithelial cells include second aggregates obtained by culturing the thawed proximal tubule epithelial cells in a state of being non-adhered onto a culture vessel which has been subjected to a cell non-adhesion process over 7 days or more;
- wherein when the thawed proximal tubule epithelial cells are cultured over 7 days or more after thawing, the gene expression level of a gene selected from the group consisting of AQP1, CD13, SGLT2, Na/K ATPase, URAT1, PEPT1, MDR1, OAT1, OCT2, OCTN2, E-cadherin, and ZO-1 in the cultured thawed proximal tubule epithelial cells are at least 10% of the gene expression level of the gene in cells of the human renal cortex.
18-21. (canceled)
A method of producing proximal tubule epithelial cells comprising:
- recovering proximal tubule epithelial cells as first aggregates;
- and
- freezing the recovered proximal tubule epithelial cells under low temperature at which the proximal tubule epithelial cells can freeze;
- thawing the frozen proximal tubule epithelial cells;
- and
- culturing the thawed proximal tubule epithelial cells;
- wherein the first aggregates are frozen while an aggregated state of the first aggregates is substantially maintained in the freezing step;
- wherein the cultured thawed proximal tubule epithelial cells include second aggregates obtained by culturing the thawed proximal tubule epithelial cells in a state of being non-adhered onto a culture vessel which has been subjected to a cell non-adhesion process over 7 days or more;
- wherein when the thawed proximal tubule epithelial cells are cultured over 7 days or more after thawing, the gene expression level of a gene selected from the group consisting of AQP1, CD13, SGLT2, Na/K ATPase, URAT1, PEPT1, MDR1, OAT1, OCT2, OCTN2, E-cadherin, and ZO-1 in the cultured thawed proximal tubule epithelial cells are at least 10% of the gene expression level of the gene in cells of the human renal cortex.
Dependencies are inferred from the claim text.