Method of producing erythrocytes

Abstract

Provided herein are methods of producing erythrocytes from hematopoietic cells, particularly hematopoietic cells from placental perfusate in combination with hematopoietic cells from umbilical cord blood, wherein the method results in accelerated expansion and differentiation of the hematopoietic cells to more efficiently produce administrable erythrocytes. Further provided herein is a bioreactor in which hematopoietic cell expansion and differentiation takes place.

What is claimed is: 
     
         1 . A method of producing erythrocytes, comprising (a) expanding a plurality of hematopoietic cells in the absence of feeder cells, optionally in contact with an immunomodulatory compound, wherein the immunomodulatory compound increases the number of hematopoietic cells compared to a plurality of hematopoietic cells not in contact with the immunomodulatory compound, to produce a first expanded hematopoietic cell population; (b) expanding the first expanded hematopoietic cell population in the presence of a plurality of feeder cells to produce a second expanded hematopoietic cell population; (c) contacting said second expanded hematopoietic cell population with one or more factors that cause differentiation of hematopoietic cells in said second expanded hematopoietic cell population into erythrocytes; and (d) isolating said erythrocytes from said second expanded hematopoietic cell population. 
     
     
         2 . The method of  claim 1 , wherein said hematopoietic cells are CD34 + . 
     
     
         3 . The method of  claim 1 , wherein said hematopoietic cells are Thy-1 + , CXCR4 + , CD133 +  or KDR + . 
     
     
         4 . The method of  claim 2 , wherein said hematopoietic cells are CD45 − . 
     
     
         5 . The method of  claim 1 , wherein said hematopoietic cells are HLA-DR − , CD71 − , CD2 − , CD3 − , CD11b − , CD11c − , CD14 − , CD16 − , CD24 − , CD56 − , CD66b −  and/or glycophorin A − . 
     
     
         6 . The method of  claim 1 , wherein said hematopoietic cells are obtained from cord blood, placental blood, peripheral blood, or bone marrow. 
     
     
         7 . The method of  claim 1 , wherein said hematopoietic cells are obtained from placental perfusate. 
     
     
         8 . The method of  claim 1 , wherein said hematopoietic cells are obtained from umbilical cord blood and placental perfusate. 
     
     
         9 . The method of  claim 6 , wherein said placental perfusate is obtained by passage of perfusion solution through only the vasculature of a placenta. 
     
     
         10 . The method of  claim 7 , wherein said placental perfusate is obtained by passage of perfusion solution through only the vasculature of a placenta. 
     
     
         11 . The method of  claim 1 , wherein said hematopoietic cells are human hematopoietic cells. 
     
     
         12 . The method of  claim 1 , wherein said feeder cells are from the same individual as said hematopoietic cells. 
     
     
         13 . The method of  claim 1 , wherein said feeder cells are from a different individual than said hematopoietic cells. 
     
     
         14 . The method of  claim 12 , wherein said feeder cells are adherent placental stem cells, bone marrow-derived mesenchymal stem cells, mesenchymal stem cells from peripheral blood, mesenchymal stem cells from cord blood, or stromal stem cells, or a combination of any of the foregoing. 
     
     
         15 . The method of  claim 12 , wherein said feeder cells are adherent placental stem cells. 
     
     
         16 . The method of  claim 15 , wherein said adherent placental stem cells are:
 CD200 +  or HLA-G + ;   CD73 + , CD105 + , and CD200 + ;   CD200 +  and OCT-4 + ;   CD73 + , CD105 +  and HLA-G + ;   CD73 +  and CD105 +  and facilitate the formation of one or more embryoid-like bodies in a population of isolated placental cells comprising said stem cells when said population is cultured under conditions that allow formation of embryoid-like bodies;   OCT-4 +  and which facilitate formation of one or more embryoid-like bodies in a population of isolated placental cells comprising said stem cell when cultured under conditions that allow formation of embryoid-like bodies;   CD10 + , CD34 − , CD105 + , and CD200 + ;   HLA-A,B,C + , CD45 − , CD133 −  and CD34 − ;   CD10 + , CD13 + , CD33 + , CD45 − , CD117 −  and CD133 − ;   CD10 − , CD33 − , CD44 + , CD45 − , and CD117 − ;   HLA A,B,C + , CD45 − , CD34 − , CD133 − ;   positive for CD10, CD13, CD38, CD44, CD90, CD105, CD200 and/or HLA-G, and/or negative for CD117;   CD200 +  and CD10 + , as determined by antibody binding, and CD117 − , as determined by both antibody binding and RT-PCR; or   CD10 + , CD29 − , CD54 + , CD200 + , HLA-G + , HLA class I +  and β-2-microglobulin + .   
     
     
         17 . The method of  claim 1 , wherein a plurality of said hematopoietic cells is blood type A, blood type O, blood type AB, blood type O; is Rh positive or Rh negative; blood type M, blood type N, blood type S, or blood type s; blood type P1; blood type Lua, blood type Lub, or blood type Lu(a); blood type K (Kell), k (cellano), Kpa, Kpb, K(a+), Kp(a−b−) or K-k-Kp(a−b−); blood type Le(a−b−), Le(a+b−) or Le(a−b+); blood type Fy a, Fy b or Fy(a−b−); or blood type Jk(a−b−), Jk(a+b−), Jk(a−b+) or Jk(a+b+). 
     
     
         18 . The method of  claim 17 , wherein the hematopoietic cells are type O, Rh positive; type O, Rh negative; type A, Rh positive; type A, Rh negative; type B, Rh positive; type B, Rh negative; type AB, Rh positive or type AB, Rh negative.