Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5811301

Title:In vitro method for producing differentiated universally compatible human blood cells
Inventors:Cameron, Robert B.; San Francisco, CA, USA
Summary:Described herein is an in vitro method of producing mature human blood cells via a process of incubating human donor pluripotent hematopoietic stem cells with a growth- or maturation-promoting polypeptide. Claims of the invention include clinically useful quantities of a single species of mature, differentiated human blood cells which are produced according to a method by which human pluripotent hematopoietic stem cells from a universal donor are incubated in a bioreactor in a growth medium which contains specific combinations of recombinant human growth and maturation promoting polypeptide factors that expand stem cell cultures and promote the maturation and differentiation of stem cells into single species of erythroid, thrombocytic or granulocytic human blood cells. Further disclosed are methods of transfecting the cells with DNA-containing vectors and genetically modifying the cells such that histocompatibility or blood group antigens with which a recipient may be incompatible are removed. Properties of the bioreactor are provided. Therapeutic applications include the use of such erythrocytes prepared in large quantities by this method as a source of iron in the form of iron-saturated hemoglobin which may be used in iron replacement therapy.
Abstract:In vitro production of clinically useful quantities of single species of mature, differentiated human blood cells is carried out 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 combinations of recombinant human growth and maturation promoting polypeptide factors that expand stem cell cultures and promote the maturation and differentiation of stem cells into single species of 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 preliminarily genetically modified so as to remove histocompatibility or blood group antigens with which a recipient may be incompatible, or the stem cells 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 also a good source of iron, in the form of iron-saturated hemoglobin, for use in iron replacement therapy.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5811301
Application Number:US1996000694478
Date Filed:07/08/1996
Date Published:22/09/1998
Assignee:None


 
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