ERYTHROID CELLS PRODUCING ADULT-TYPE BETA-HEMOGLOBIN GENERATED FROM HUMAN EMBRYONIC STEM CELLS
Methods and compositions of erythroid cells that produce adult β-hemoglobin, generated by culturing CD31+, CD31+/CD34+ or CD34+ cells from embryonic stem cells under serum-free culture conditions.
1 - 15 . (canceled)
16 . A population of human erythroid cells derived from pluripotent stem cell culture, wherein the erythroid cells produce adult-type β-hemoglobin but not embryonic c-hemoglobin, and wherein essentially all of the cells within the population are erythroid cells.
17 . The cell population of claim 16 , wherein essentially all live cells are nucleated erythroid cells.
18 . The cell population of claim 16 , wherein the population is essentially free of lymphocytes.
19 . The cell population of claim 16 , wherein the population contains less than 0.1% leukocytes.
20 . The cell population of claim 16 , wherein the population is CD31 − , CD34 − , CD71 + and CD 235a + .
21 . A preparation of CD31+/CD34+/CD43+ hematopoietic progenitor cells, wherein the cells have a potential to differentiate into erythroid cells that produce adult-type β-hemoglobin but not embryonic c-hemoglobin.
22 . The preparation of claim 21 , wherein the cells are the result of 6 to 7 days (±10%) of a co-culture comprising OP9 cells.
23 . An isolated population of human hematopoietic progenitor cells, comprising 10% to 20% CD31 + /CD43 + hematopoietic progenitors, up to 60% CD31 + /CD43 − /KDR bright endothelial cells and less than 15% CD34 + /CD43 − /KDR− mesenchymal cells, wherein the cells have a potential to differentiate into erythroid cells that produce adult-type β-hemoglobin but not embryonic ζ-hemoglobin.
24 . An isolated population of human hematopoietic progenitor cells, comprising 10% to 20% CD34 + /CD43 + hematopoietic progenitors, up to 60% CD34 + /CD43 − /KDR bright endothelial cells and less than 15% CD34 + /CD43 − /KDR − mesenchymal cells, wherein the cells have a potential to differentiate into erythroid cells that produce adult-type β-hemoglobin but not embryonic ζ-hemoglobin.
25 . An isolated population of human hematopoietic progenitor cells, wherein the cells are derived from human pluripotent stem cells and are at least one of CD31+ and CD34+ and wherein the cells have a potential to differentiate into erythroid cells that produce adult-type β-hemoglobin but not embryonic ζ-hemoglobin.
26 . The isolated population of hematopoietic progenitor cells of claim 25 , wherein the population comprises CD31+/CD34+/CD43+hematopoietic progenitor cells.
27 . The isolated population of human hematopoietic progenitor cells of claim 25 , wherein the cells are greater than 95% pure.
28 . A population of human erythroid cells derived from the hematopoietic progenitor cells of claim 25 .
29 . The population of human erythroid cells of claim 28 , wherein the erythroid cells produce adult-type β-hemoglobin but not embryonic ζ-hemoglobin, and wherein essentially all of the cells within the population are erythroid cells.
30 . The population of human erythroid cells of claim 28 , wherein essentially all live cells are nucleated erythroid cells.
31 . The population of human erythroid cells of claim 28 , wherein the population is essentially free of lymphocytes.
32 . The population of human erythroid cells of claim 28 , wherein the population contains less than 0.1% leukocytes.
33 . The population of human erythroid cells of claim 28 , wherein the population is CD31 − , CD34 − , CD71 + and CD235 + .