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 ce
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.
대표청구항▼
1. A method of producing erythrocytes, comprising (a) expanding a population of isolated human CD34+ hematopoietic stem cells from a plurality of human hematopoietic stem cells in a serum-free medium in contact with one or more factors in the absence of feeder cells, wherein said CD34+ hematopoietic
1. A method of producing erythrocytes, comprising (a) expanding a population of isolated human CD34+ hematopoietic stem cells from a plurality of human hematopoietic stem cells in a serum-free medium in contact with one or more factors in the absence of feeder cells, wherein said CD34+ hematopoietic stem cells are in contact with pomalidomide or lenalidomide for a time and in an amount sufficient for the pomalidomide or lenalidomide to increase the number of CD34+ hematopoietic stem cells compared to a population of human CD34+ hematopoietic stem cells not in contact with the pomalidomide or lenalidomide, to produce a first expanded CD34+ hematopoietic stem cell population; wherein said factors comprise SCF, Flt-3 and IL-3,(b) expanding the first expanded CD34+ hematopoietic stem cell population in a medium in contact with one or more factors in the presence of a plurality of feeder cells, wherein said feeder cells are adherent placental stem cells, to produce a second expanded human CD34+ hematopoietic stem cell population; wherein said second population of human CD34+ hematopoietic stem cells differentiate into erythrocytes during said expanding, wherein said factors comprise SCF, Epo and Insulin-like growth factor I (IGF-1), and(d) isolating said erythrocytes from said second expanded hematopoietic stem cell population. 2. The method of claim 1, wherein said hematopoietic stem cells are obtained from a source selected from the group consisting of umbilical cord blood, placental blood, peripheral blood, and bone marrow. 3. The method of claim 1, wherein said hematopoietic stem cells are obtained from placental perfusate. 4. The method of claim 1, wherein said hematopoietic stem cells are obtained from umbilical cord blood and placental perfusate. 5. The method of claim 4, wherein said placental perfusate is obtained by passage of perfusion solution through only the vasculature of a placenta. 6. The method of claim 3, wherein said placental perfusate is obtained by passage of perfusion solution through only the vasculature of a placenta. 7. The method of claim 1, wherein said feeder cells are from the same individual as said hematopoietic stem cells. 8. The method of claim 1, wherein said feeder cells are from a different individual than said hematopoietic stem cells. 9. The method of claim 1, wherein said adherent placental stem cells are selected from the group consisting of: (a) CD200+ or HLA-G+;(b) CD73+, CD105+, and CD200+;(c) CD200+ and OCT-4+;(d) CD73+, CD105+ and HLA-G+;(e) CD10+, CD34−, CD105+, and CD200+;(f) HLA-A,B,C+, CD45−, CD133− and CD34−;(g) CD10+, CD13+, CD33+, CD45−, CD117− and CD133−;(h) CD10−, CD33−, CD44+, CD45−, and CD117−;(i) HLA A,B,C+, CD45−, CD34−, CD133−;(j) positive for CD10, CD13, CD38, CD44, CD90, CD105, and are one or more of CD200 positive, HLA-G positive, and CD117 negative;(k) CD200+ and CD10+, as determined by antibody binding, and CD117−, as determined by both antibody binding and RT-PCR; and(l) CD10+, CD29−, CD54+, CD200+, HLA-G+, HLA class I+ and β-2-microglobulin+. 10. The method of claim 1, wherein a plurality of said hematopoietic stem cells has a blood type selected from the group consisting of blood type A, blood type B, blood type AB, and blood type O, wherein a plurality of said hematopoietic stem cells is Rh positive or Rh negative, and wherein a plurality of said hematopoietic stem cells optionally has a blood type selected from the group consisting of-blood type M, blood type N, blood type S, blood type s, blood type P1, blood type Lua, blood type Lub, blood type Lu(a), blood type K (Kell), blood type k (cellano), blood type Kpa, blood type Kpb, blood type K(a+), blood type Kp(a−b−), blood type K− k− Kp(a−b−), blood type Le(a−b−), blood type Le(a+b−), blood type Le(a−b+), blood type Fy a, blood type Fy b, blood type Fy(a−b−), blood type Jk(a−b−), blood type Jk(a+b−), blood type Jk(a−b+), and blood type Jk(a+b+). 11. The method of claim 10, wherein the hematopoietic stem 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.
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이 특허에 인용된 특허 (111)
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