Methods, compositions, and growth and differentiation factors for insulin-producing cells

• US 20040259244A1
• Filed: 03/15/2004
• Published: 12/23/2004
• Est. Priority Date: 05/28/2002
• Status: Abandoned Application

• Current Assignee: Becton, Dickinson & Co, Novocell, Inc. (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Becton, Dickinson & Co
• Inventors: Presnell, Sharon C., Haaland, Perry D., Heidaran, Mohammad A., Coutts, Margaret, McIntyre, Catherine Anne, Latta, Paul Presley, Scharp, David William
• Application Number:
  US10/800,813
• Publication Number:
  US 20040259244A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• 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: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/46 Amines, e.g. putrescine
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/113 Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/315 : Prolactin
• CPC Class Codes: C12N 2501/335 : Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/34 : Calcitonin; Calcitonin-gene related peptide [CGRO]; Amylin
• CPC Class Codes: C12N 2501/345 : Gastrin; Cholecystokinins [CCK]
• CPC Class Codes: C12N 2501/35 : Vasoactive intestinal peptide [VIP]; Pituitary adenylate cyclase activating polypeptide [PACAP]
• CPC Class Codes: C12N 2501/37 : Parathyroid hormone [PTH]
• CPC Class Codes: C12N 2501/39 : Steroid hormones
• CPC Class Codes: C12N 2501/392 : Sexual steroids
• CPC Class Codes: C12N 2501/41 : Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/83 : Tachykinins, e.g. substance P
• CPC Class Codes: C12N 2501/85 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/22 : from pancreatic cells
• CPC Class Codes: C12N 5/0037 : Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

A method of converting differentiated non-hormone producing pancreatic cells into differentiated hormone producing cells is disclosed. The method comprises two steps: first, culturing cells under conditions which convert differentiated non-hormone producing cells into stem cells; and second, culturing stem cells under conditions which provide for differentiating stem cells into hormone-producing cells. The invention defines growth and differentiation factors that are presented to the stem cells to result in their differentiation into hormone-producing cells, especially insulin-producing cells. The invention provides a new source of large quantities of hormone producing cells such as insulin-producing cells that are riot currently available for therapeutic uses such as the treatment of diabetes.

Claims

1. A method of converting differentiated non-hormone producing pancreatic cells into differentiated hormone-producing cells, comprising:

a) culturing said differentiated non-hormone producing pancreatic cells in a first cell culture system with a first cell culture medium comprising a basal medium, with or without serum, and with or without growth factors;

under conditions which provide for converting said differentiated non-hormone producing pancreatic cells into stem cells; and

b) culturing said stem cells in a second cell culture system with a second cell culture medium comprising at least one compound selected from Group A, wherein the compounds of Group A are selected from the group consisting of;

Betacellulin, Activin A, BMP-2, TGF-β

SRII, DMSO, Sonic Hedgehog, Laminin, Met-Enkephalin, DMF, and Cholera Toxin A;

and at least one compound selected from Group B, wherein the compounds of Group B are selected from the group consisting of;

Activin A, Atrial Natriuretic Peptide, Betacellulin, Bone Morphogenic Protein (BMP-2), Bone Morphogenic Protein (BMP-4), C natriuretic peptide (CNP), Caerulein, Calcitonin Gene Related Peptide (CGRP-ax), Cholecystokinin (CCK8-amide), Cholecystokinin octapeptide (CCK8-sulfated), Cholera Toxin B Subunit, Corticosterone (Reichstein'"'"'s substance H), Dexamethasone, DIF-1, Differanisole A, Dimethylsulfoxide (DMSO), EGF, Endothelin 1, Exendin 4, FGF acidic, FGF2, FGF7, FGFb, Gastrin I, Gastrin Releasing Peptide (GRP), Glucagon-Like Peptide 1 (GLP-1), Glucose, Growth Hormone, Hepatocyte Growth Factor (HGF), IGF-1, IGF-2, Insulin, KGF, Lactogen, Laminin, Leu-Enkephalin, Leukemia Inhibitory Factor (LIF), Met-Enkephalin, n Butyric Acid, Nerve Growth Factor (β

-NGF), Nicotinamide, n-n-dimethylformamide (DMF), Parathyroid Hormone Related Peptide (Pth II RP), PDGF AA+PDGF BB MIX, PIGF (Placental GF), Progesterone, Prolactin, Putrescine Dihydrochloride Gamma-Irradiated Cell Culture, REG1, Retinoic Acid, Selenium, Selenious Acid, Sonic Hedgehog, Soybean Trypsin Inhibitor, Substance P, Superoxide Dismutase (SOD), TGF-α

, TGF-β

sRII, TGF-β

1, transferrin, Triiodothyronine (T3), Trolox, Vasoactive Intestinal Peptide (VIP), VEGF, Vitamin A, and Vitamin E;

under conditions which provide for differentiating said stem cells into hormone-producing cells.

2. The method of claim 1, wherein the second cell culture medium comprises at least two compounds selected from Group A and at least two compounds selected from Group B.

3. The method of claim 1, wherein the second cell culture medium comprises at least three compounds selected from Group A and at least three compounds selected from Group B.

4. The method of claim 1, wherein the second cell culture medium comprises at least four compounds selected from Group A and at least four compounds selected from Group B.

5. The method of claim 1, wherein the second cell culture medium comprises at least five compounds selected from Group A and at least five compounds selected from Group B.

6. The method of claim 1, wherein the second cell culture medium comprises at least six compounds selected from Group A and at least six compounds selected from Group B.

7. A method of culturing stem cells into differentiated hormone-producing cells, comprising culturing the stem cells in a cell culture system with a cell culture medium whereby said stem cells are differentiated into hormone-producing cells wherein said culture medium comprises basal medium without serum and at least one compound selected from Group A wherein the compounds of Group A are selected from the group consisting of:

Betacellulin, Activin A, BMP-2, TGF-β

SR11, DMSO, Sonic Hedgehog, Laminin, Met-Enkephalin, DMF, and Cholera Toxin A;

and at least one compound selected from Group B, wherein the compounds of Group B are selected from the group consisting of;

Activin A, Atrial Natriuretic Peptide, Betacellulin, Bone Morphogenic Protein (BMP-2), Bone Morphogenic Protein (BMP-4), C natriuretic peptide (CNP), Caerulein, Calcitonin Gene Related Peptide (CGRP-α

), Cholecystokinin (CCK8-amide), Cholecystokinin octapeptide (CCK8-sulfated), Cholera Toxin B Subunit, Corticosterone (Reichstein'"'"'s substance H), Dexamethasone, DIF-1, Differanisole A, Dimethylsulfoxide (DMSO), EGF, Endothelin 1, Exendin 4, FGF acidic, FGF2, FGF7, FGFb, Gastrin I, Gastrin Releasing Peptide (GRP), Glucagon-Like Peptide 1 (GLP-1), Glucose, Growth Hormone, Hepatocyte Growth Factor (HGF), IGF-1, IGF-2, Insulin, KGF, Lactogen, Laminin, Leu-Enkephalin, Leukemia Inhibitory Factor (LIF), Met-Enkephalin, n Butyric Acid, Nerve Growth Factor (β

-NGF), Nicotinamide, n-n-dimethylformamide (DMF), Parathyroid Hormone Related Peptide (Pth II RP), PDGF AA+PDGF BB MIX, PIGF (Placental GF), Progesterone, Prolactin, Putrescine Dihydrochloride Gamma-Irradiated Cell Culture, RE G1, Retinoic Acid, Selenium, Selenious Acid, Sonic Hedgehog, Soybean Trypsin Inhibitor, Substance P, Superoxide Dismutase (SOD), TGF-α

, TGF-β

sRII, TGF-β

1, transferrin, Triiodothyronine (T3), Trolox, Vasoactive Intestinal Peptide (VIP), VEGF, Vitamin A, and Vitamin E.

8. The method of claim 7, wherein the cell culture medium comprises at least two compounds selected from Group A and at least two compounds selected from Group B.

9. The method of claim 7, wherein the cell culture medium comprises at least three compounds selected from Group A and at least three compounds selected from Group B.

10. The method of claim 7, wherein the cell culture medium comprises at least four compounds selected from Group A and at least four compounds selected from Group B.

11. The method of claim 7, wherein the cell culture medium comprises at least five compounds selected from Group A and at least five compounds selected from Group B.

12. The method of claim 7, wherein the cell culture medium comprises at least six compounds selected from Group A and at least six compounds selected from Group B.