Conditioned cell culture medium compositions and methods of use

• US 8,138,147 B2
• Filed: 01/06/2009
• Issued: 03/20/2012
• Est. Priority Date: 05/14/1999
• Status: Expired due to Fees

• Current Assignee: Allergan Incorporated (Abbvie Incorporated)
• Original Assignee: Skinmedica Incorporated (Abbvie Incorporated)
• Inventors: Naughton, Gail K., Horwitz, David L., Applegate, Mark A., Zeltinger, Joan, Mansbridge, Jonathan N., Kern, Andreas, Landeen, Lee K., Ratcliffe, Anthony, Pinney, R. Emmett
• Primary Examiner(s): Gamett, Daniel C
• Application Number:
  US12/349,451
• Publication Number:
  US 20090123503A1
• Time in Patent Office: 1,169 Days
• Field of Search: None
• US Class Current: 514/8.1
• CPC Class Codes: A23L 33/10 using additives addition of substantially indigestible substances A23L33/21
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 2800/86 Products or compounds obtained by genetic engineering
• CPC Class Codes: A61K 2800/91 Injection
• CPC Class Codes: A61K 35/33 Fibroblasts
• CPC Class Codes: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61K 35/36 Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells islets of Langerhans A61K35/39
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 38/18 Growth factors; Growth regulators
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 38/39 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 8/02 characterised by special physical form
• CPC Class Codes: A61K 8/0208 Tissues; Wipes; Patches
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61K 8/65 Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
• CPC Class Codes: A61K 8/985 Skin or skin outgrowth, e.g. hair, nails
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2430/36 : for embolization or occlusion, e.g. vaso-occlusive compositions or devices
• CPC Class Codes: A61L 31/044 : Collagen
• CPC Class Codes: A61L 31/047 : Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
• CPC Class Codes: A61L 31/16 : Biologically active materials, e.g. therapeutic substances A61L31/047 takes precedence
• CPC Class Codes: A61P 1/00 : Drugs for disorders of the alimentary tract or the digestive system
• 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 1/18 : for pancreatic disorders, e.g. pancreatic enzymes
• 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 17/14 : for baldness or alopecia
• CPC Class Codes: A61P 17/16 : Emollients or protectives, e.g. against radiation
• CPC Class Codes: A61P 17/18 : Antioxidants, e.g. antiradicals preparations for protection against sunlight A61Q17/00
• CPC Class Codes: A61P 19/00 : Drugs for skeletal disorders
• CPC Class Codes: A61P 19/08 : for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 39/06 : Free radical scavengers or antioxidants
• CPC Class Codes: A61P 41/00 : Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61Q 19/00 : Preparations for care of the skin
• CPC Class Codes: A61Q 19/08 : Anti-ageing preparations
• CPC Class Codes: A61Q 7/00 : Preparations for affecting hair growth
• CPC Class Codes: C12N 2502/1323 : Adult fibroblasts
• 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: C12N 5/0625 : Epidermal cells, skin cells; Cells of the oral mucosa
• CPC Class Codes: C12N 5/0629 : Keratinocytes; Whole skin
• CPC Class Codes: C12N 5/0656 : Adult fibroblasts
• CPC Class Codes: C12N 5/069 : Vascular Endothelial cells
• CPC Class Codes: C12P 21/00 : Preparation of peptides or proteins single cell protein C12N1/00

Abstract

Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. The medium may be conditioned by stromal cells, parenchymal cells, mesenchymal stem cells, liver reserve cells, neural stem cells, pancreatic stem cells and/or embryonic stem cells. Additionally, the cells may be genetically modified. A three-dimensional tissue construct is preferred. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder. Additionally, the medium is formulated with a pharmaceutically acceptable carrier as a vehicle for internal administration, applied directly to a food item or product, formulated with a salve or ointment for topical applications, or, for example, made into or added to surgical glue to accelerate healing of sutures following invasive procedures. Also, the medium may be further processed to concentrate or reduce one or more factors or components contained within the medium.

Claims

1. A method of making a composition, comprising:

(a) forming a conditioned medium comprising culturing fibroblasts in a nutrient medium sufficient to meet the nutritional needs required to grow the cells in vitro wherein the conditioned medium comprises at least 3.2 ng/ml vascular endothelial growth factors (VEGF);

(b) removing the conditioned medium from the cultured fibroblasts; and

(c) combining the conditioned medium with an acceptable carrier to form the composition.

2. The method of claim 1, wherein the conditioned medium obtained from said cultured fibroblasts further comprises keratinocyte growth factor (KGF).

3. The method of claim 2, wherein said conditioned medium contains at least 1.67 ng/ml KGF.

4. The method of claim 1, wherein culturing comprises a continuous flow system.

5. The method of claim 1, wherein culturing comprises a bioreactor system.

6. The method of claim 1, wherein the cultured fibroblasts are suspended or floated in the nutrient medium.

7. The method of claim 1, further comprising processing said conditioned medium, wherein the conditioned medium in step (b) is concentrated, filtered, purified, or a combination thereof prior to combining with an acceptable carrier in step (c).

8. The method of claim 1, wherein approaching physiological levels is performed by concentrating said conditioned medium.

9. The method of claim 1, wherein said conditioned medium further comprises at least 2.3 ng/ml G-CSF.

10. The method of claim 1, wherein said conditioned medium further comprises at least 0.9 ng/ml IL-8.

11. The method of claim 1, wherein said conditioned medium further comprises at least 0.8 ng/ml TGF-β

.

12. A cosmeceutical composition comprising:

cell culture media, conditioned by;

(a) culturing fibroblasts in a nutrient medium sufficient to meet the nutritional needs required to grow the cells in vitro wherein the conditioned medium comprises at least 3.2 ng/ml vascular endothelial growth factors (VEGF), so that a conditioned medium is formed;

(b) removing the conditioned medium from the cultured cells; and

(c) combining the conditioned medium with an acceptable carrier to form the composition.

13. The cosmeceutical composition of claim 12, further comprising processing said conditioned medium wherein the conditioned medium is concentrated, filtered, purified, or a combination thereof prior to combining with an acceptable carrier in step (c).

14. The cosmeceutical composition of claim 12, wherein approaching physiological levels is performed by concentrating said conditioned medium.

15. The cosmeceutical composition of claim 12, wherein the conditioned medium obtained from said cultured fibroblasts comprises VEGF and keratinocyte growth factor (KGF).

16. The cosmeceutical composition of claim 12, wherein said composition is an injectable composition or a topical composition.

17. The cosmeceutical composition of claim 16, wherein said topical composition is an ointment, a cream, a hydrogel or a lotion.

18. The conditioned medium of claim 15, wherein the conditioned medium contains at least 1.67 ng/ml KGF.

19. A method of making a composition, comprising:

(a) forming a conditioned medium comprising culturing fibroblasts in a nutrient medium sufficient to meet the nutritional needs required to grow the cells in vitro, wherein said cultured fibroblasts form an extracellular matrix and secrete a desired level of extracellular products so that a conditioned medium is formed, wherein at least 3.2 ng/ml of vascular endothelial growth factor (VEGF) is secreted into the conditioned medium;

(b) removing the conditioned medium from the cultured fibroblasts; and

(c) combining the conditioned medium with an acceptable carrier to form the composition.

20. The method of claim 19, wherein the conditioned medium obtained from said cultured fibroblasts further comprises keratinocyte growth factor (KGF).

21. The method of claim 20, wherein said conditioned medium contains at least 1.67 ng/ml KGF.

22. The method of claim 19, wherein culturing comprises a continuous flow system.

23. The method of claim 19, wherein culturing comprises a bioreactor system.

24. The method of claim 19, wherein the cultured fibroblasts are suspended or floated in the nutrient medium.

25. The method of claim 19, further comprising processing said conditioned medium, wherein the conditioned medium in step (b) is concentrated, filtered, purified, or a combination thereof prior to combining with an acceptable carrier in step (c).