Compositions comprising human embryonic stem cells and their derivatives, methods of use, and methods of preparation
First Claim
1. A method for testing the effect of a compound on hES cells, derivatives of hES cells, or combinations thereof, wherein the derivatives of hES cells are selected from the group consisting of hematopoietic stem cell progenitors, neuronal stem cell progenitors, mesenchymal stem cell progenitors, insulin producing stem cell progenitors, hepatocytic stem cell progenitors, cardiac stem cell progenitors, epithelial stem cell progenitors, and combinations thereof, and wherein the hES cells and derivatives of hES cells are obtained by culturing in media that is free from feeder cells, animal products, growth factors, leukemia inhibiting factor, fibroblast growth factor, vitamin supplements, membrane associated steel factor, soluble steel factor, and conditioned media, and further by method (1),wherein method (1) comprises expanding the cells free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor, and conditioned media, comprising:
- (i) introducing the cells in a cell culture medium consisting of minimal essential medium, a progestin, and a β
-human chorionic gonadotropin (β
hCG) agonist; and
(ii) incubating the cells at a temperature of about 34°
C. to about 38°
C. in an environment of about 3.5% to about 6% carbon dioxide for about 12 hours to about 48 hours; and
in the presence of the compound and determining the effect of the compound on the cells.
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Abstract
The present invention relates to a pharmaceutical composition comprising of preparations of human embryonic stem (hES) cells and their derivatives and methods for their transplantation into the human body, wherein transplantation results in the clinical reversal of symptoms, cure, stabilization or arrest of degeneration of a wide variety of presently incurable and terminal medical conditions, diseases and disorders. The invention further relates to novel processes of preparing novel stem cell lines which are free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor and conditioned media. This invention further relates to the isolation, culture, maintenance, expansion, differentiation, storage, and preservation of such stem cells.
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Citations
4 Claims
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1. A method for testing the effect of a compound on hES cells, derivatives of hES cells, or combinations thereof, wherein the derivatives of hES cells are selected from the group consisting of hematopoietic stem cell progenitors, neuronal stem cell progenitors, mesenchymal stem cell progenitors, insulin producing stem cell progenitors, hepatocytic stem cell progenitors, cardiac stem cell progenitors, epithelial stem cell progenitors, and combinations thereof, and wherein the hES cells and derivatives of hES cells are obtained by culturing in media that is free from feeder cells, animal products, growth factors, leukemia inhibiting factor, fibroblast growth factor, vitamin supplements, membrane associated steel factor, soluble steel factor, and conditioned media, and further by method (1),
wherein method (1) comprises expanding the cells free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor, and conditioned media, comprising: -
(i) introducing the cells in a cell culture medium consisting of minimal essential medium, a progestin, and a β
-human chorionic gonadotropin (β
hCG) agonist; and(ii) incubating the cells at a temperature of about 34°
C. to about 38°
C. in an environment of about 3.5% to about 6% carbon dioxide for about 12 hours to about 48 hours; andin the presence of the compound and determining the effect of the compound on the cells. - View Dependent Claims (2)
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3. A method for testing the effect of a compound on hES cells, derivatives of hES cells, or combinations thereof, wherein the derivatives of hES cells are selected from the group consisting of hematopoietic stem cell progenitors, neuronal stem cell progenitors, mesenchymal stem cell progenitors, insulin producing stem cell progenitors, hepatocytic stem cell progenitors, cardiac stem cell progenitors, epithelial stem cell progenitors, and combinations thereof, and wherein the hES cells and derivatives of hES cells are obtained by culturing in media that is free from feeder cells, animal products, growth factors, leukemia inhibiting factor, fibroblast growth factor, vitamin supplements, membrane associated steel factor, soluble steel factor, and conditioned media, and further by method (2),
wherein method (2) comprises partially differentiating the cells free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor, and conditioned media, comprising: -
(i) introducing the cells, wherein the cells are obtained by expansion, in a cell culture medium consisting of minimal essential medium, wherein the cell culture medium does not contain a progestin or a β
-human chorionic gonadotrophic (β
hCG) agonist; and(ii) incubating the cells at a temperature of about 34°
C. to about 38°
C. in an environment of about 3.5% to about 6% carbon dioxide for about 12 hours to about 48 hours in a substantially aerobic environment,in the presence of the compound and determining the effect of the compound on the cells. - View Dependent Claims (4)
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Specification