Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5599705

Title:In vitro method for producing differentiated universally compatible mature human blood cells
Inventors:Cameron, Robert B.; Agoura Hill, CA, USA
Summary:Described herein is a method for the in vitro production of mature human blood cells via a process of incubating the human donor pluripotent hematopoietic stem cells with a growth or maturation promoting polypeptide. The invention relates to the generation of significant quantities of mature, differentiated human blood cells by a method in which human pluripotent hematopoietic stem cells that have been obtained from a universal donor are incubated in a bioreactor with a growth medium which contains specific recombinant human growth and maturation promoting polypeptide factors in combinations that expand stem cell cultures and which promote the maturation and differentiation of stem cells into erythroid, thrombocytic or granulocytic human blood cells. Further disclosed are methods of harvesting the mature cells after they have been grown and allowed to mature in the presence of polypeptides such as SCGF, Interleukins 1,3,4,5,6, and 11, GM-CSF, M-CSF, G-CSF and EPO. Also provided are methods for the modification of stem cells such that their histo-incompatibility or blood group antigens may be removed or genetically altered by transfection with appropriate DNA-containing vectors prior to their addition to the bioreactor. Claims of the invention include the use of such erythrocytes prepared in large quantities by this method as a good source of iron such as iron-saturated hemoglobin. Therapeutic applications are provided for use in iron replacement therapy.
Abstract:In vitro production of clinically useful quantities of mature, differentiated human blood cells by a method in which human pluripotent hematopoietic stem cells, preferably from a universal donor, are incubated in a bioreactor in a growth medium also containing specific recombinant human growth and maturation promoting polypeptide factors in combinations that expand stem cell cultures and promote the maturation and differention of stem cells into erythroid, thrombocytic or granulocytic human blood cells, and harvesting the mature cells. The growth and maturation promoting polypeptides employed include SCGF, Interleukins 1,3,4,5,6, and 11, GM-CSF, M-CSF, G-CSF and EPO. Stem cells may be modified so as to remove histocompatibility and/or blood group antigens, or may be genetically altered by transfection with appropriate DNA-containing vectors, prior to addition to the bioreactor. Erythrocytes prepared in large quantities by this method are a good source of iron, as iron-saturated hemoglobin, for use in iron replacement therapy.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5599705
Application Number:US1993000152389
Date Filed:16/11/1993
Date Published:04/02/1997
Assignee:None


 
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