Compositions comprising human embryonic stem cells and their derivatives, methods of use, and methods of preparation

• US 9,482,660 B2
• Filed: 10/24/2013
• Issued: 11/01/2016
• Est. Priority Date: 03/07/2006
• Status: Active Grant

• Current Assignee: Geeta Shroff
• Original Assignee: Geeta Shroff
• Inventors: Shroff, Geeta
• Primary Examiner(s): Lankford, Blaine
• Application Number:
  US14/062,512
• Publication Number:
  US 20140051110A1
• Time in Patent Office: 1,104 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/48 Reproductive organs
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/00 : General protective or antinoxious agents
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2501/31 : Pituitary sex hormones, e.g. follicle-stimulating hormone [FSH], luteinising hormone [LH]; Chorionic gonadotropins
• CPC Class Codes: C12N 2501/392 : Sexual steroids
• CPC Class Codes: C12N 5/0603 : Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0606 : Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: G01N 33/5044 : involving specific cell types

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.

Claims

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.

2. The process of claim 1, further comprising:

(e) storing the preparation at about −

15°

C. to about −

72°

C., and

(f) thawing the stored preparation naturally before transplantation,

wherein the viability of the cells is at least 40% on thawing.

5. A process of preparing the ready to use preparation of claim 1, further comprising testing the preparation for any contamination prior to transplantation.

6. The process of claim 2, wherein the viability of the cells is at least 50% upon thawing.

7. The process of claim 2, wherein the viability of the cells is at least 60% upon thawing.

8. The process of claim 2, wherein the viability of the cells is at least 70% upon thawing.

9. The process of claim 2, wherein the viability of the cells is at least 80% upon thawing.

10. The process of claim 2, wherein the ready to use preparation further comprises an antimicrobial agent, an antibacterial agent, a hormonal product, or an other pharmaceutical agent.

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:

(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.

4. The process of claim 3, further comprising:

(g) storing the preparation at about −

15°

C. to about −

72°

C., and

(h) thawing the stored preparation naturally before transplantation,

wherein the viability of the cells is at least 40% on thawing.

11. A process of preparing the ready to use preparation of claim 3, further comprising testing the preparation for any contamination prior to transplantation.

12. The process of claim 4, wherein the viability of the cells is at least 50% upon thawing.

13. The process of claim 4, wherein the viability of the cells is at least 60% upon thawing.

14. The process of claim 4, wherein the viability of the cells is at least 70% upon thawing.

15. The process of claim 4, wherein the viability of the cells is at least 80% upon thawing.

16. The process of claim 4, wherein the ready to use preparation further comprises an antimicrobial agent, an antibacterial agent, a hormonal product, or an other pharmaceutical agent.