Compositions comprising human embryonic stem cells and their derivatives, methods of use, and methods of preparation
First Claim
1. A process of preparing a ready to use preparation of hES cells, derivatives of hES cells, or combinations thereof for human transplantation, 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, comprising:
- (a) introducing the cells in a cell culture medium consisting of minimal essential medium, a progestin, and a β
-human chorionic gonadotropin (β
hCG) agonist;
(b) expanding the cells obtained in (a) 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;
(c) centrifuging the cells obtained in (b) to obtain a pellet, and(d) suspending the pellet in a biocompatible solution.
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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
16 Claims
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1. A process of preparing a ready to use preparation of hES cells, derivatives of hES cells, or combinations thereof for human transplantation, 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, comprising:
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(a) introducing the cells in a cell culture medium consisting of minimal essential medium, a progestin, and a β
-human chorionic gonadotropin (β
hCG) agonist;(b) expanding the cells obtained in (a) 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;(c) centrifuging the cells obtained in (b) to obtain a pellet, and (d) suspending the pellet in a biocompatible solution. - View Dependent Claims (2, 5, 6, 7, 8, 9, 10)
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3. A process of preparing a ready to use preparation of hES cells, derivatives of hES cells, or combinations thereof for human transplantation, 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, comprising:
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(a) introducing the cells in a cell culture medium consisting of minimal essential medium, a progestin, and a β
-human chorionic gonadotropin (β
hCG) agonist;(b) expanding the cells obtained in (a) 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;(c) introducing the cells obtained in (b) to a cell culture medium consisting of minimal essential medium; (d) incubating the cells obtained in (c) 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, wherein the cells are incubated in a biocompatible container under substantially aerobic conditions, and wherein the cells differentiate upon proliferation;(e) centrifuging said stem cells to obtain a pellet, and (f) suspending the pellet in a biocompatible solution. - View Dependent Claims (4, 11, 12, 13, 14, 15, 16)
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Specification