Method for producing endothelial cells
Inventors
Yamamoto, Yuki • Enoki, Tatsuji • Tosaka, Yasuhiro • Yamaguchi, Yoko • Mineno, Junichi
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11225643-B2
Publication Date
2022-01-18
Expiration Date
Abstract
The present invention relates to a method for producing endothelial cells, including carrying out: (a) inducing a population of mesoderm-lineage cells containing endothelial progenitor cells from pluripotent stem cells without forming an embryoid body; and (b) culturing the population of mesoderm-lineage cells containing endothelial progenitor cells in the presence of RepSox, in this order. According to the present invention, endothelial cells with high quality can be efficiently produced from pluripotent stem cells. The endothelial cells obtained by the method of the present invention are useful for the production of, for example, a myocardial sheet, and expected to be utilized in the treatment of a heart disease. A myocardial sheet can be produced by mixing the endothelial cells obtained by the method of the present invention with myocardial cells and mural cells and culturing the cells.
Core Innovation
A method is disclosed for producing endothelial cells from human pluripotent stem cells. The method induces a population of mesoderm-lineage cells comprising endothelial progenitor cells from human pluripotent stem cells without forming an embryoid body, and then obtains isolated endothelial progenitor cells by isolating the endothelial progenitor cells from the population of mesoderm-lineage cells.
The isolated endothelial progenitor cells are then cultured in the presence of RepSox. During step (a), the human pluripotent stem cells are sequentially cultured in a first medium comprising activin A, a second medium comprising bone morphogenetic protein 4, and a third medium comprising vascular endothelial growth factor.
The disclosed approach includes isolation of endothelial progenitor cells that are kinase insert domain receptor-positive and comparison against control cells without RepSox. The method includes an outcome constraint of an increase in CD31-positive cells at day 19 compared to control cells.
Claims Coverage
One independent claim is present. It contains three main inventive feature groups: EB-free induction of mesoderm-lineage cells to obtain isolated endothelial progenitor cells, sequential induction media for the pluripotent stem cells using activin A, bone morphogenetic protein 4, and vascular endothelial growth factor, and RepSox culture of isolated endothelial progenitor cells with an outcome constraint of increased CD31-positive cells at day 19 versus control.
Embryoid body-free mesoderm-lineage induction and isolation of endothelial progenitors
Inducing a population of mesoderm-lineage cells comprising endothelial progenitor cells from human pluripotent stem cells without forming an embryoid body, and isolating the endothelial progenitor cells from the population of mesoderm-lineage cells to obtain isolated endothelial progenitor cells.
Sequential induction media using activin A, bone morphogenetic protein 4, and vascular endothelial growth factor
In step (a), the human pluripotent stem cells are sequentially cultured in a first medium comprising activin A, a second medium comprising bone morphogenetic protein 4, and a third medium comprising vascular endothelial growth factor.
RepSox culturing with CD31-positive increase at day 19
Culturing the isolated endothelial progenitor cells in the presence of RepSox, wherein there is an increase in CD31-positive cells at day 19 compared to control cells.
Across the independent claim, endothelial production is anchored by EB-free induction of mesoderm-lineage cells to generate isolated endothelial progenitors, sequential pluripotent induction using activin A, bone morphogenetic protein 4, and vascular endothelial growth factor, and subsequent RepSox culturing, with the key measured outcome being increased CD31-positive cells at day 19 versus control.
Stated Advantages
Increase in CD31-positive cells at day 19 compared to control cells.
Documented Applications
Not explicitly described in patent.
Interested in licensing this patent?