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US 2026/0234561 A1Published application
Method for producing platelets
AssigneeDewcell, Inc.
InventorsCHI HWA KIM, CHI YEON HWANG
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Abstract
A method for producing platelets includes culturing pluripotent stem cells to obtain a culture medium containing hematopoietic stem cells, obtaining a suspended cell population from the culture medium when the cells in the culture medium that do not express CD41a constitute at least 15% of the total cells, and differentiating the suspended cell population into megakaryocytes, whereby the method enables the production of high-quality platelets with a high yield.
Claims15
1:
- A method for producing platelets, the method comprising:
- culturing pluripotent stem cells to obtain a culture containing hematopoietic stem cells;
- obtaining a floating cell population from the culture when the number of CD41a+ cells among the cells contained in the culture is at least 15% of the total number of cells;
- and
- differentiating the floating cell population into megakaryocytes in a first medium.
2 more elements
2:
- The method for producing platelets according to claim 1, further comprising culturing and maturing the megakaryocytes in a second medium containing thrombopoietin.
3:
- The method for producing platelets according to claim 1, wherein the culturing of the pluripotent stem cells comprises:
- culturing the pluripotent stem cells in a third medium containing a GSK3 inhibitor;
- culturing the cells cultured in the third medium in a fourth medium containing vascular endothelial growth factor and basic fibroblast growth factor;
- and
- culturing the cells cultured in the fourth medium in a fifth medium containing vascular endothelial growth factor;
- basic fibroblast growth factor;
- and a transforming growth factor beta signaling inhibitor.
4 more elements
4:
- The method for producing platelets according to claim 1, wherein the floating cell population has a CD45+ cell count of 50% or less of the total cell count.
5:
- The method for producing platelets according to claim 1, wherein the floating cell population has a CD34+ cell count of 45% or more of the total cell count.
6:
- The method for producing platelets according to claim 1, wherein the floating cell population has a CD41a− cell count of 85% or less of the total cell count.
7:
- The method for producing platelets according to claim 1, further comprising (9)-seeding the pluripotent stem cells on the bottom of a culture vessel at 2,000 to 20,000 cells/cm2.
8:
- The method for producing platelets according to claim 1, wherein the culture further comprises hematopoietic progenitor cells and megakaryocyte progenitor cells.
9:
- The method for producing platelets according to claim 1, wherein the first medium comprises thrombopoietin, stem cell factor, interleukin-3, and interleukin-6.
10:
- The method for producing platelets according to claim 1, wherein, in the obtaining of the floating cell population, the floating cell population is obtained from the culture when the number of CD41a+ cells among the cells contained in the culture is 40 to 85% of the total number of cells.
11:
- The method for producing platelets according to claim 1, wherein, in the obtaining of the floating cell population, the floating cell population is obtained from the culture when the number of CD34+ cells among the cells contained in the culture is 45 to 80% of the total number of cells.
12:
- The method for producing platelets according to claim 1, wherein, in the obtaining of the floating cell population, the floating cell population is obtained from the culture when the number of CD45+ cells among the cells contained in the culture is 1 to 50% of the total number of cells.
13:
- The method for producing platelets according to claim 1, wherein the pluripotent stem cells are human induced pluripotent stem cells.
14:
- The method for producing platelets according to claim 4, wherein the floating cell population has a proportion of CD45+ cells lower than a proportion of CD41a+ cells.
15:
- A method for producing a blood product, the method comprising mixing platelets produced by the method of claim 1 with other blood components.