Conditioned cell culture medium compositions and methods of use
원문보기
IPC분류정보
국가/구분
United States(US) Patent
공개
국제특허분류(IPC7판)
A61K-035/12
C12N-005/08
A61K-048/00
출원번호
US-0538380
(2006-10-03)
공개번호
US-0077232
(2007-04-05)
발명자
/ 주소
Naughton,Gail, K.
Horwitz,David, L.
Applegate,Mark, A.
Zeltinger,Joan
Mansbridge,Jonathan, N.
Kern,Andreas
Landeen,Lee, K.
Ratcliffe,Anthony
Pinney,R., Emmett
출원인 / 주소
SkinMedica, Inc.
대리인 / 주소
WILSON SONSINI GOODRICH & ROSATI
인용정보
피인용 횟수 :
0인용 특허 :
0
초록▼
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 condi
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.
대표청구항▼
1. A pharmaceutical composition comprising conditioned cell culture medium, said medium comprising medium which has previously supported the growth of eukaryotic cells cultured in three-dimensions, and a pharmaceutical carrier. 2. The pharmaceutical composition of claim 1 wherein the conditioned
1. A pharmaceutical composition comprising conditioned cell culture medium, said medium comprising medium which has previously supported the growth of eukaryotic cells cultured in three-dimensions, and a pharmaceutical carrier. 2. The pharmaceutical composition of claim 1 wherein the conditioned cell culture medium is in the form of a liquid, solid, lyophilized, powder, gel or film. 3. The pharmaceutical composition of claim 1 wherein the composition comprises the reduction or enrichment of one or more extracellular products derived from the conditioned media by protein separation techniques including immunoaffinity chromatography, gel chromatography, ion exchange, metal chelate affinity chromatography, HPLC purification and hydrophobic interaction chromatography. 4. The pharmaceutical composition of claim 3 wherein the extracellular product(s) is an extracellular matrix protein. 5. The pharmaceutical composition of claim 3 wherein the extracellular product(s) is a growth factor, anti-inflammatory mediator, enzyme, cytokine, hormone, clotting factor, regulatory factor, angiogenic factor, or anti-angiogenic factor. 6. The pharmaceutical composition of claim 1 wherein the cells cultured in three-dimensions comprise stromal cells and form a three-dimensional stromal tissue. 7. The three-dimensional stromal tissue of claim 6 comprising stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three-dimensional structure. 8. The pharmaceutical composition of claim 7 wherein the framework structure comprises a mesh, sponge or polymerized hydrogel. 9. The pharmaceutical composition of claim 8 wherein the stromal cells comprise mesenchymal stem cells, liver reserve cells, neural stem cells, pancreatic stem cells, fibroblast-like cells, fibroblast cells, chondro-progenitor cells, chondrocytes, embryonic stem cells, endothelial cells, pericytes, macrophages, monocytes, plasma cells, mast cells or any combination thereof. 10. The pharmaceutical composition of claim 9 wherein the stromal cells are human. 11. The pharmaceutical composition of claim 9 or 10 wherein the stromal cells comprise genetically engineered cells. 12. The pharmaceutical composition of claim 6 wherein parenchymal cells are cultured on the three-dimensional stromal tissue to form a tissue-specific three-dimensional tissue construct. 13. The pharmaceutical composition of claim 12 wherein the parenchymal cells comprise skin, hepatic, kidney, neuronal, pancreatic, intestinal, genitourinary, vascular, spleen, bone, bone marrow, or mucosal cells. 14. The pharmaceutical composition of claim 13 wherein the parenchymal cells comprise genetically engineered cells. 15. A food substance with enhanced nutritional content comprising conditioned cell culture medium, said medium comprising medium which has previously supported the growth of eukaryotic cells together with a separate food item for consumption by a mammal. 16. The food substance of claim 15 wherein the separate food item is suitable for human consumption. 17. A nutritional supplement comprising conditioned cell culture medium, said medium comprising medium which has previously supported the growth of eukaryotic cells, together with a carrier suitable for human consumption and where said carrier is in the form of a liquid, tablet or capsule. 18. A method of making a pharmaceutical composition comprising: (a) culturing eukaroytic cells in three-dimensions in a cell culture medium sufficient to meet the nutritional needs required to grow the cells in vitro; (b) culturing the cells in vitro until the cell culture medium contains a desired level of extracellular products so that a conditioned medium is formed; (c) removing the conditioned medium from the cells used to condition the medium; and (d) combining the conditioned medium with a pharmaceutical carrier. 19. The method of claim 18 wherein the conditioned medium is recovered from a continuous flow system. 20. The method of claim 18 or 19 wherein the conditioned medium is cultured in an aseptic environment. 21. The method claim 18 wherein the conditioned cell medium is processed into the state of a liquid, solid, lyophilized, powder, gel or film. 22. The method of claim 18 wherein the conditioned medium is further processed to concentrate or reduce one or more products contained in the medium. 23. The method of claim 22 wherein the composition comprises the reduction or enrichment of one or more extracellular products derived from the conditioned media by protein separation techniques including immunoaffinity chromatography, gel chromatography, ion exchange, metal chelate affinity chromatography, HPLC purification and hydrophobic interaction chromatography. 24. The method of claim 22 wherein the extracellular product(s) is an extracellular matrix protein. 25. The method of claim 22 wherein the extracellular product(s) is a growth factor, anti-inflammatory mediator, enzyme, cytokine, hormone, antigen, antibody, clotting factor, regulatory factor, angiogenic factor, or anti-angiogenic factor. 26. The method of claim 18 wherein the cells cultured in three-dimensions comprise stromal cells and form a three-dimensional stromal tissue. 27. The method of claim 26 comprising stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed into a three-dimensional structure. 28. The method of claim 27 wherein the three-dimensional framework comprises a mesh, sponge or a polymerized hydrogel. 29. The method of claim 26 or 27 wherein the stromal cells comprise mesenchymal stem cells, liver reserve cells, neural stem cells, pancreatic stem cells, fibroblast-like cells, fibroblast cells, chondro-progenitor cells, chondrocytes, embryonic stem cells, endothelial cells, pericytes, macrophages, monocytes, plasma cells, mast cells or any combination thereof. 30. The method of claim 26 wherein the stromal cells comprise genetically engineered cells. 31. The method of claim 30 wherein the genetically engineered cells are transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed and secreted into the conditioned medium. 32. The method of claim 26 wherein parenchymal cells are cultured on the three-dimensional stromal tissue to form a tissue-specific three-dimensional tissue culture. 33. The method of claim 32 wherein the parenchymal cells are skin, hepatic, kidney, neuronal, pancreatic, intestinal, genitourinary, kidney, spleen, bone, bone marrow, or mucosal cells. 34. The method of claim 33 wherein the parenchymal cells comprise genetically engineered cells. 35. The method of claim 34 wherein the genetically engineered cells are transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed and secreted into the conditioned medium. 36. A method for improved wound or burn healing comprising administering to a patient in need of wound or burn healing treatment a composition comprising cell culture medium, said medium comprising medium which has previously supported the growth of eukaryotic cells cultured in three-dimensions, together with a separate therapeutic component so that the patient exhibits a reduction in the amount of traumatized tissue or scar tissue and further exhibits improved regeneration of healthy tissue at the wound site. 37. The method of claim 36 wherein the wound is a vascular wound. 38. The method of claim 36 wherein the wound is to the brain or spinal cord. 39. The method of claim 36 wherein the wound is to the skin, liver, kidney, pancreas, intestines, spleen, genitourinary tract, bone, bone marrow or mucosal tissue. 40. The method of claim 36 wherein the component is a bandage. 41. The method of claim 36 wherein the component is an ointment or cream and the composition is applied topically. 42. The method of claim 36 wherein component is a surgical glue or wound filler. 43. The method of claim 36 wherein the component is a suture or implant and is coated with the conditioned medium prior to use. 44. The method of claim 36 wherein the component is a pharmaceutical carrier so that the conditioned medium is in the form of an injectable, tablet or capsule. 45. The method of claim 36 wherein the composition further comprises an antibiotic, anti-inflammatory, antiviral, antifungal, hormone, antitumor, analgesic, anesthetic, or any combination thereof. 46. The method of claim 36 wherein the cells cultured in three-dimensions contain stromal cells and form a three-dimensional stromal tissue. 47. The method of claim 46 wherein the three-dimensional stromal tissue comprises stromal cells attached to and substantially enveloping framework composed of a biocompatible, non-living material formed into a three-dimensional structure. 48. The method of claim 47 wherein the framework structure comprises a mesh, sponge or hydrogel. 49. The method of claim 46, wherein the stromal cells are mesenchymal stem cells, fibroblast-like cells, fibroblast cells, chondro-progenitor cells, chondrocytes, embryonic stem cells, endothelial cells, pericytes, macrophages, monocytes, plasma cells, mast cells or any combination thereof. 50. The method of claim 49 wherein the stromal cells are genetically engineered cells. 51. The method of claim 46 wherein parenchymal cells are cultured on the three-dimensional stromal tissue to form a tissue-specific three-dimensional tissue construct. 52. The method of claim 51 wherein the parenchymal cells comprise skin, hepatic, kidney, neuronal, pancreatic, intestinal, genitourinary, vascular, spleen, bone, bone marrow or mucosal cells. 53. The method of claim 52 wherein the parenchymal cells are genetically engineered cells. 54. The method of claim 53 wherein the genetically engineered cells are transfected with an exogenous gene under the control of an expression element so that the exogenous gene product is expressed and secreted into the conditioned medium. 55. A method for the correction of a cosmetic defect comprising administering to a person a composition comprising conditioned cell culture medium, said medium comprising medium which has previously supported the growth of eukaryotic cells cultured in three-dimensions, together with a component useful for cosmetic purposes, so that the person exhibits a cosmetic improvement. 56-57. (canceled) 58. A method for the stimulation of hair growth comprising topically administering to a person a composition comprising conditioned cell culture medium, said medium comprising medium which has previously the growth of eukaryotic cells cultured in three dimensions, together with a topical carrier, so that the person exhibits an improved stimulation of hair growth. 59. (canceled) 60. A method for isolating collagen, comprising: (a) precipitating procollagen from medium conditioned by a three-dimensional culture at high salt conditions under neutral pH conditions; (b) removing the propeptides under acidic conditions so that the triple helix of the collagen remains intact; and (c) precipitating the collagen at high salt concentrations.
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