Erythroid cells producing adult-type beta-hemoglobin generated from human embryonic stem cells

Abstract

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 + .