WO2002091955A1 - Rein artificiel presentant la fonction de metabolisme de proteine et son procede de fabrication - Google Patents
Rein artificiel presentant la fonction de metabolisme de proteine et son procede de fabrication Download PDFInfo
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- WO2002091955A1 WO2002091955A1 PCT/JP2002/004778 JP0204778W WO02091955A1 WO 2002091955 A1 WO2002091955 A1 WO 2002091955A1 JP 0204778 W JP0204778 W JP 0204778W WO 02091955 A1 WO02091955 A1 WO 02091955A1
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- WIPO (PCT)
- Prior art keywords
- artificial kidney
- megalin
- growth factor
- cells
- sponge sheet
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/14—Dialysis systems; Artificial kidneys; Blood oxygenators ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3813—Epithelial cells, e.g. keratinocytes, urothelial cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3839—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by the site of application in the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/26—Materials or treatment for tissue regeneration for kidney reconstruction
Definitions
- the present invention provides a sponge sheet into which a hydrogel containing a growth factor that promotes angiogenesis has been injected, and an artificial kidney having a protein metabolizing function characterized by comprising a cell expressing megalin on its surface, And a method for manufacturing the same.
- hemodialysis therapy The weakness of hemodialysis therapy is that it cannot compensate for the glomerular filtration protein metabolism function of proximal tubular cells. Therefore, low molecular weight proteins originally metabolized by the cells accumulate in the body of hemodialysis patients and act as uretoxin proteins to cause various disease states.
- a typical example is dialysis amyloidosis caused by accumulation of J32-microglobulin (] 32-m), which causes osteoarticular disorders and organ failure.
- glycation-modifying protein, AGE also accumulates and is involved in arteriosclerosis and organ damage.
- AGE glycation-modifying protein
- the present invention provides an in-vivo artificial kidney having a protein metabolism function and a method for producing the same, in order to solve the above-mentioned problems.
- the present invention also provides an artificial kidney having a protein metabolizing function by utilizing the function of megalin, which binds to low-molecular-weight proteins and hormones to be taken into cells.
- the present invention relates to a sponge sheet into which a hydrogel containing a growth factor that promotes angiogenesis has been injected, and megalin, an endcytosis receptor that plays a central role in the protein metabolism of proximal tubular cells.
- the present invention relates to an in-vivo artificial kidney having a protein metabolizing function, which comprises a cell expressed on the surface as a component.
- the present invention provides an in-vivo artificial kidney having a protein metabolizing function, characterized by using a sponge sheet injected with a hydrogel containing a growth factor for promoting angiogenesis, and cells expressing megalin on the surface. It relates to a manufacturing method. Further, the present invention provides a step of injecting a hydrogel containing a growth factor that promotes angiogenesis into a sponge sheet; a step of implanting the sponge sheet subcutaneously; and a step of superimposing megalin on the implanted sponge sheet. The present invention relates to a method for producing the above-mentioned artificial kidney, which comprises a step of injecting cells expressed in the artificial kidney.
- the growth factor that promotes angiogenesis is basic fibroblast growth factor (bFGF); bFGF is human bFGF produced using genetic engineering techniques; and the open-mouth gel is a gelatin gel;
- the gelatin gel is composed mainly of gelatin having an isoelectric point of 4.5 to 5.5; the sponge sheet is made of collagen; the cells expressing megalin on the surface are transformed into human proximal tubule epithelial cells and Z or by gene transfer. Human proximal tubular epithelial cells with high expression of megalin; megalin expression is due to genetic manipulation.
- the in-vivo artificial kidney having a protein metabolizing function of the present invention is characterized by comprising, as constituent components, a sponge sheet into which a hydrogel containing a growth factor for promoting angiogenesis has been injected, and cells expressing megalin on the surface. I do.
- growth factors that promote angiogenesis used in the present invention include basic fibroblast growth factor (bFGF; basic fibroblast growth factor), acidic fibroblast growth factor (a FGF), and epidermal growth factor.
- bFGF basic fibroblast growth factor
- a FGF acidic fibroblast growth factor
- epidermal growth factor epidermal growth factor
- EGF epidermall growth factor
- TGF- ⁇ transforming growth factor-
- VEGF vascular endothelial cell growth factor
- HGF hepatocyte growth factor
- the growth factors that promote angiogenesis may be used, or a plurality of them may be used in combination. Furthermore, it is also possible to use a combination of factors having other biological activities.
- bFGF vascular endothelial growth factor
- PDGF-BB vascular endothelial growth factor
- bFGF has been widely accumulated as an angiogenesis promoting factor, and is one of the more preferably used growth factors.
- the bFGF used in the present invention includes those extracted from organs or tissues such as pituitary gland, brain, kidney, adrenal gland, placenta, bone matrix, cartilage, endothelial cells, fibroblasts, and genes such as genetic modification. It includes those produced using engineering techniques and those that act as fibroblast growth factors, such as modified forms thereof. Among them, human bFGF produced using genetic engineering techniques is particularly preferred from the viewpoint of quality and supply stability.
- the above-mentioned modified product is a product obtained by adding an amino acid to the amino acid sequence of bFGF obtained by extraction or genetic engineering techniques, or a part of the amino acid is substituted with another amino acid. And those in which some of the amino acids have been deleted. In the present invention It is possible to use these FGFs alone or as a mixture.
- the hydrogel used in the present invention is used for sustaining the angiogenic effect by gradually releasing the contained growth factor that promotes angiogenesis into the vicinity thereof.
- Raw materials of the hydrogel used in the present invention include, for example, polysaccharides such as cellulose, dextran, agarose, pullulan, starch, hyaluronic acid, alginic acid, chitin, chitosan, and hydroxyethyl cellulose and carboxymethyl cellulose.
- polysaccharide derivatives polyamino acids such as polyaspartic acid, polydaltamic acid, and polylysine; polypeptides such as gelatin, collagen, fibrin, and gluten; polyvinyl alcohol, polyacrylamide, polyvinylinolepyrrolidone, and poly. (2-Hydroxyethynolemetharylate), Polyvinyl methyl ether, Poly (N-Bulacetamide), Poly (ataryl acid), Poly (isobutylene-maleic acid), Poly (2-acrylylamide-2-Methyl Pansu sulfonic acid), poly acryloxy propane sulfonic acid, polyvinyl sulfonic acid, Po
- V Metal-oic quishethyl quaternized ammonium chloride
- polybutylpyridine poly (N, N-dimethyl-N— (2-metararyloyloxetyl) —N- (3-sulfopropyl) ammonium internal salt) And the like.
- Synthetic polymers having a hydrophilic group in the side chain such as synthetic polymers having a main chain itself such as polyethylene dalicol, polydioxolan, and polyethyleneimine.
- these materials can be used alone or in appropriate combination of two or more, and further modified by other compounds.
- polysaccharides such as cellulose, hyaluronic acid, alginic acid, chitin, chitosan, polyamino acids such as polyaspartic acid, polyglutamic acid, and polylysine
- polypeptides such as gelatin, collagen, and fipurin
- gelatin is more preferably used in view of easiness of processing, maintenance of biological activity of a growth factor to be contained, and degradation rate in vivo (slow release of growth factor).
- Gelatin can be obtained by subjecting collagen contained in a living body to an appropriate pretreatment with an acid, alkali, or the like, followed by heat extraction using warm water.
- gelatin can be used without any particular limitation as long as it is generally available. Can be.
- examples thereof include alkali-treated gelatin having an isoelectric point of 4.5 to 5.5, and acid-treated gelatin having an isoelectric point of 8 to 9.
- the isoelectric point of bFGF is about 9 and it is positively charged in a neutral aqueous solution.
- Alkali-treated gelatin having a point of 4.5 to 5.5 is preferably used.
- gelatin not only one kind of gelatin, but also different kinds of raw materials or different kinds of gelatins having different physical properties such as solubility, molecular weight and isoelectric point can be used. Further, other additives may be contained. For example, as disclosed in JP-A-8-325160, b Addition of a polyone compound or the like for imparting sustained release of FGF, etc. Can also be used. When bFGF is used as an angiogenesis-promoting factor, even when the above-mentioned hydrogel other than gelatin is used, a hydrogel having a low isoelectric point is preferable from the viewpoint of the retention of bFGF. The duration of the angiogenic effect can be varied depending on the biodegradability and the water content of the hydrogel used.
- the water content of the hydrogel is preferably 80% or more, more preferably 85 to 99%, and more preferably 90 to 98% when swollen in water, from the viewpoint of the sustained release of growth factors that promote angiogenesis. % Is more preferred.
- the water content w of the hydrogel can be determined as follows.
- V D and V w As a method for measuring V D and V w , for example, there is a method in which the operation of dropping a measuring cylinder from a height of about 5 cm is repeated to read the volume when the volume no longer changes.
- V w because it may take time to settle in the hydrogel in water, by reading the volume at which the dropping operation was carried out about 10 times, the volume when allowed to stand for 30 minutes longer changed
- the wet weight for example, when a hydrogel is taken on a glass filter and suctioned using an aspirator, and a weight loss curve indicating the relationship between suction time and weight is created, the slope of the curve is gentle.
- the drying method is not particularly limited as long as the method can achieve a constant weight. However, normal pressure drying at a temperature of 105 ° C. or more can be easily employed without requiring any special equipment. However, when decomposition or the like occurs due to drying at this temperature, a vacuum drying method at a low temperature such as freeze-drying can be adopted.
- the hydrogel used in the present invention is obtained by water insolubilization by crosslinking, but depending on the origin and properties of the material, heat treatment, irradiation with ultraviolet light or ⁇ -ray, curing agent A cross-linking method such as a reaction with
- the curing agent is not particularly limited.
- an inorganic compound such as a salt containing a polyvalent metal ion such as aluminum or ferric iron; an aldehyde such as dartalaldehyde or formalin; Carpimides such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, 1-cyclohexyl 3- (2-morpholinoethyl) carbodiimide-methyl ⁇ -toluenesulfonate, epichlorohydride
- examples thereof include epoxy compounds such as phosphorus and butanediol diglycidyl ether, isocyanates such as hexamethylene diisosinate, and organic compounds such as acid anhydride.
- Examples of the shape of the hydrogel used in the present invention include a column, a sheet, a disk, a sphere, a particle, an irregular shape, and the like.From the viewpoint of easy injection into a sponge sheet, a sphere, a particle, Amorphous ones are preferably used.
- the size of the hydrogel is preferably from 10 ⁇ m to 200 ⁇ , more preferably from 20 ⁇ to 150 ⁇ .
- the sponge sheet used in the present invention is used for fixing a hydrogel containing a growth factor that promotes angiogenesis, and for holding cells expressing megalin on the surface. Is a porous sheet.
- polyesters such as polyglycolic acid, polylactic acid, polycaprolactone, polydioxanone, polyhydroxyacetic acid, polyhydroxyvaleric acid, polytrimethylene carbonate, poly Bioabsorbable materials such as (a-cyanoacrylate), polyurethane and the like.
- these materials may be used alone or in combination of two or more, and may be used after being modified with another compound.
- the raw materials for the sponge sheet collagen, polyglycolic acid, polylactic acid, and the like are preferable from the viewpoint of a track record as a biocompatible material.
- a collagen sponge sheet is more preferably used because of its good cell engraftment.
- the sponge sheet used in the present invention has a porous structure.However, from the viewpoint of the ease of invasion of new blood vessels and the supply of oxygen and nutrients to cells to be injected, the sponge sheet has an all-porous structure having communicating pores. Is preferred.
- the average pore diameter is preferably from 60 to 400 im, more preferably from 70 to 150 / zm.
- Megalin is an endocytic receptor that plays a central role in the protein metabolism of proximal tubule cells whose cDNA has been successfully cloned in rats by the present inventors, Saito et al., And whose primary structure has been analyzed. (Proc. Natl. Acad. Sc USA 91, p. 9725-9729 (1994)). Thereafter, Hja1mG et al. Cloned human megalin / gp330 and analyzed the entire primary structure (Eur. J. Biochem. 239, ppl32-137, 1996)).
- Human megalin is estimated to be 4655-aa (molecular weight 519, 636), N-terminal (25-aa), extracellular region (4398-aa), transmembrane domain (23-aa), C-terminal cytoplasmic region ( 209-aa), and the extracellular region has three types of cysteine-rich regions and is known to belong to the LDL receptor gene family.
- Megalin is an enzyme or enzyme inhibitor such as PA I-1, PA 1-1 perokinase, PA I-1 t-PA, plow mouth kinase, lipoprotein lipase, aprotune, vitamin D binding protein, retinol binding protein, etc.
- Vitamin binding proteins apoli Apolipoproteins such as poprotein B and apolipoprotein E, basic polypeptides such as aminoglycosides and polymixin B, parathyroid hormone, insulin, 2_m, low epidermal growth factor, prolatatin, lysozyme, cytochrome C, etc. It is a multifunctional endocytic receptor that has been reported to bind to high molecular weight proteins such as hormones.
- megalin is expressed in humans in addition to renal proximal tubular epithelial cells, parathyroid (parathyroid) cells, epithelial cell layer in placental intervillous space, epididymal epithelial cells, type III It has been confirmed that it is expressed on the surface of alveolar epithelial cells, breast epithelial cells, thyroid follicle cells, and ciliary epithelial cells. That is, these cells and the like are mentioned as megalin-expressing cells.
- proximal tubular epithelial cells collected from autologous renal kidney are particularly preferably used.
- the cells expressing megalin on the surface thereof used in the present invention include, in addition to the above-mentioned autologous or other autologous megalin-expressing cells, autologous or autologous megalin-expressing cells to which genes have been transfected to express megalin, and autologous cells.
- known physical methods, chemical methods, biological methods, and the like can be used.
- Examples of the method of inducing differentiation include a method of providing a differentiation-inducing substance, a method of co-culturing with other cells, and a method of changing the environment such as temperature, pressure, pH, and osmotic pressure.
- autologous cells are preferable from the viewpoint of the exclusion of cells implanted in the body. Further, human proximal tubular epithelial cells and / or human proximal tubular epithelial cells in which megalin is highly expressed by gene transfer are more preferable.
- a macrophage-like cell with high phagocytic ability derived from autologous peripheral blood is preferably used because of its simplicity in technique.
- megalin is preferably by genetic manipulation.
- the artificial kidney of the present invention is produced using a sponge sheet into which a hydrogel containing a growth factor that promotes angiogenesis has been injected, and cells that have expressed megalin on the surface.
- the method for producing an artificial kidney includes the steps of: injecting a hydrogel containing a growth factor that promotes angiogenesis into a sponge sheet; implanting the sponge sheet subcutaneously; and implanting the sponge. Including a step of injecting cells expressing megalin on the surface of the sheet.
- a method for producing a hydrogel a known method can be used without any particular limitation.
- a method using physical aggregation between molecules by hydrogen bond, ionic bond, coordination bond, or the like, a method of chemically cross-linking using a cross-linking agent, a method of cross-linking by irradiation with light or radiation, and the like can be mentioned.
- the method for molding (formulation) of the hydrogel is not particularly limited.
- an emulsion method, a melt molding method, a spray drying method and the like can be used.
- any method for incorporating a growth factor that promotes angiogenesis into the hydrogel any method can be used without particular limitation as long as the biological activity of the growth factor that promotes angiogenesis is not significantly impaired.
- a method of drying a hydrogel after producing a hydrogel and absorbing a solution containing a growth factor into the hydrogel or a method of impregnating a solution containing a growth factor without drying after producing a hydrogel, and a method of producing a hydrogel Occasionally, there is a method in which a growth factor is present in the stock solution.
- the hydrogel containing a growth factor that promotes angiogenesis can be injected into a sponge sheet using, for example, a syringe.
- the sponge sheet is used by implanting it in a living body such as subcutaneous in order to fix a hydrogel containing a growth factor that promotes angiogenesis and to retain cells expressing megalin on the surface.
- a hydrogel containing a growth factor that promotes neogenesis may be implanted into a sponge sheet and then implanted, or the sponge sheet may be implanted and then injected into the hydrogel.
- the in-vivo artificial kidney of the present invention is obtained by injecting a hydrogel containing a growth factor that promotes angiogenesis and injecting cells expressing megalin into a sponge sheet implanted in a living body. It can be manufactured by the following.
- Cells expressing megalin on the surface include, for example, syringes, microcapillaries, etc. Can be injected into the sponge sheet.
- FIG. 1 is a graph showing the incorporation of 125 I-labeled] 32-m into tissues in Examples and Comparative Examples. The notation in the figure is as follows.
- FIG. 2 shows the results of SDS-polyacrylamide gel electrophoresis analysis of [ 125 I-labeled] 32-m incorporated into transplanted cells.
- the samples in each lane in the figure are as follows.
- Lane 1 125 I-labeled / 32-m used for intraperitoneal administration
- Lane 2 homogenate of transplanted cell mass collected after the test
- Lane 3 blood of cell-transplanted mice collected after the test.
- FIG. 3 is a graph showing the results of radioisotope counting during precipitation of tricloacetic acid (TCA) in blood in Examples and Comparative Examples. The notation in the figure is as follows. c o n t r o l g r o u p;
- b FGF-containing gelatin gel particles were prepared according to the method of Tabata et al. That is, 1 OmL of a 10 wt% alkali-treated gelatin (isoelectric point 5.0) aqueous solution (manufactured by Nitta Gelatin) preheated to 40 ° C and 25 / zL of a 25 wt% glutaraldehyde aqueous solution were mixed. The solution was dropped into 375 mL of olive oil under stirring at 40 ° C. and 425 rpm to form a w / o emulsion. After that, the gelatin was chemically cross-linked by continuously stirring at 25 ° C for 24 hours.
- the obtained particles were collected by centrifugation (4 ° C., 3000 rpm, 5 minutes), and further washed by centrifugation in acetone for 5 times.
- the unreacted aldehyde groups of dartartaldehyde are sealed by keeping the washed particles in lO OmL of 100 mM glycine aqueous solution containing 0.1 wt% Tween 80 for 1 hour at 37 ° C. did.
- the obtained crosslinked gelatin gel particles were washed by centrifugation twice in distilled water, freeze-dried, and sterilized with ethylene oxide gas. The water content of the gelatin gel particles after swelling in water at 37 ° C for 24 hours was 95%.
- the particle diameter (diameter) measured with an optical microscope was 60 to 130 ⁇ .
- a 0.3% atelocollagen hydrochloride solution (pH 3.0) was stirred for 60 minutes at 1800 to 2000 rpm using a refrigerated homogenizer.
- the foamed solution was poured into a mold, rapidly frozen at 40 ° C, freeze-dried for 48 hours, and further dried under reduced pressure at 105 ° C for 24 hours.
- phosphate-buffered saline (pH 7. Washed in 4) and immersed in a 15% aqueous ethanol solution. Freeze quickly at 135 ° C, freeze-dry (48 hours), sterilize with ethylene oxide gas,
- the bFGF-containing gelatin gel particles obtained in the above (1) are suspended in 0.15 mL of PBS, and the IX 1 X0.5 cm collagen sponge sheet obtained in the above (2) is used for lmL. It was injected with a syringe (23 G needle). The sponge sheet was implanted subcutaneously in the dorsal side of a 5-week-old female nude mouse (BALBZcA Jc1 nu; CLEA Japan) under anesthesia by inhalation of getyl ether.
- L2 cells Dulbecco's Modified Eagle medium (LI FE TECHNOLOG IES, GI) supplemented with 10% (vol / vol) neonatal sera was added to a sponge sheet whose angiogenesis was pathologically confirmed.
- the cells (L2 cells) derived from rat yolk sac epithelial carcinoma cells, which express megalin on the surface, were cultured in PBS (BCO BR L, Rockvi 11e, MD, USA). 0.15 mL of the cell suspension (10 VmL) was injected with a 1 mL syringe (23 G needle) under mouse anesthesia. Two weeks after the cell injection, both kidneys of mice whose longest cell mass had grown to 2 cm or more were surgically removed from the dorsal side under anesthesia to obtain renal failure. No metastasis of the transplanted L2 cells was observed.
- the cell suspension was not injected, and only PBS was used as collagen.
- the procedure was the same as in the example, except that it was injected into the sheet.
- FIG. 1 is a graph showing the incorporation of 125 I-labeled 2-m into tissues in Examples and Comparative Examples.
- the vertical axis shows 125 I counts per unit weight of tissue.
- 125 I counts in transplanted cells were significantly (*: p ⁇ 0.05) higher than those in heart, lung, liver and skeletal muscle at any time measured.
- heart, lung, 125 1 count at month interference organ and skeletal muscle there was no significant difference in any of the time measured in Example and Comparative Example.
- transplanted cell mass was treated with ULTRA TURRAX (IKAKA) in Laemmli sample buffer supplemented with 2% -mercaptoethanol.
- FIG. 2 is a diagram showing the results of SDS-polyacrylamide gel electrophoresis analysis of 125 m-labeled 2-m incorporated into transplanted cells.
- the transplanted cells is seen to contain a 125 I-labeled J3 2-m monomer and its degradation product.
- ⁇ 2-m chain was found in larger amounts than in the transplanted cells.
- FIG. 3 is a graph showing the results of radioisotope counting during TCA precipitation of blood in Examples and Comparative Examples. Radioisotope counting in TCA precipitation of blood, 1 25 after I-labeled 2-tn administered 6 and 14 hours, significant in Example than in Comparative Example (*
- the artificial kidney having a protein metabolism function of the present invention can exert its function in the body, and is useful for improving QOL of patients with renal failure.
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Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP02769619A EP1388327B1 (en) | 2001-05-17 | 2002-05-17 | Artificial kidney having function of metabolizing protein and method of constructing the same |
CA2445785A CA2445785C (en) | 2001-05-17 | 2002-05-17 | Artificial kidney having function of metabolizing protein and method of constructing the same |
JP2002588876A JP4181877B2 (ja) | 2001-05-17 | 2002-05-17 | 蛋白代謝機能を有する人工腎臓およびその作製方法 |
US10/478,170 US20040235161A1 (en) | 2001-05-17 | 2002-05-17 | Artificial kidney having function of metabolizing protein and method of constructing the same |
KR1020037014699A KR100864648B1 (ko) | 2001-05-17 | 2002-05-17 | 단백질 대사 기능을 갖는 인공 신장 및 그 제작 방법 |
HK04106757A HK1064024A1 (en) | 2001-05-17 | 2004-09-07 | Combination for constructing intra-corporeal artificial kidney having function of metabolizing protein |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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JP2001-147699 | 2001-05-17 | ||
JP2001147699 | 2001-05-17 |
Publications (1)
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WO2002091955A1 true WO2002091955A1 (fr) | 2002-11-21 |
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Application Number | Title | Priority Date | Filing Date |
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PCT/JP2002/004778 WO2002091955A1 (fr) | 2001-05-17 | 2002-05-17 | Rein artificiel presentant la fonction de metabolisme de proteine et son procede de fabrication |
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Country | Link |
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US (1) | US20040235161A1 (ja) |
EP (1) | EP1388327B1 (ja) |
JP (1) | JP4181877B2 (ja) |
KR (1) | KR100864648B1 (ja) |
CN (1) | CN100335015C (ja) |
CA (1) | CA2445785C (ja) |
HK (1) | HK1064024A1 (ja) |
TW (1) | TWI221421B (ja) |
WO (1) | WO2002091955A1 (ja) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2005094912A1 (ja) * | 2004-03-31 | 2005-10-13 | Cardio Incorporated | 血管新生作用が強化された移植片 |
JP2006042795A (ja) * | 2004-06-30 | 2006-02-16 | Hokkaido Univ | 細胞培養用基材、その製造方法及び細胞培養方法 |
WO2007029677A1 (ja) * | 2005-09-09 | 2007-03-15 | Gunze Limited | 組織再生用基材 |
WO2007119563A1 (ja) | 2006-03-28 | 2007-10-25 | Niigata University | ヒトメガリンの測定方法 |
Families Citing this family (12)
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EP2075019B1 (en) * | 2006-10-17 | 2012-03-07 | Tokai University Educational System | Bioartificial renal tubule |
DE102007039871A1 (de) * | 2007-08-21 | 2009-02-26 | Friedrich-Baur-Gmbh | Weichgewebe-Implantat mit antibakterieller Wirkung |
NZ590122A (en) * | 2008-07-02 | 2012-02-24 | Otsuka Pharma Co Ltd | Artificial kidney precursor comprising a non-human mammalian metanephros and process for the production thereof |
JP5424702B2 (ja) | 2009-04-27 | 2014-02-26 | 国立大学法人 新潟大学 | 尿中ヒトメガリンを測定することを含む腎疾患検出方法 |
WO2010126055A1 (ja) | 2009-04-27 | 2010-11-04 | 国立大学法人新潟大学 | 腎障害の検出用マーカーとしての尿中メガリンの使用 |
WO2011040599A1 (ja) | 2009-10-02 | 2011-04-07 | シャープ株式会社 | 血管状態モニタリング装置およびモニタリング方法 |
EP2485755A4 (en) * | 2009-10-06 | 2014-04-02 | Agency Science Tech & Res | BMP-7 ADMINISTRATION AND METHODS OF USE |
US9173604B2 (en) * | 2010-03-19 | 2015-11-03 | Sharp Kabushiki Kaisha | Measurement device, measurement method, measurement result processing device, measurement system, measurement result processing method, control program, and recording medium |
CN103025762A (zh) * | 2010-06-22 | 2013-04-03 | Jjk医疗有限公司 | 新介质、装置和方法 |
EP3006559B1 (en) * | 2013-05-31 | 2019-11-06 | iHeart Japan Corporation | Layered cell sheet incorporating hydrogel |
EP3437660A4 (en) * | 2016-03-28 | 2019-05-01 | Fujifilm Corporation | PREPARATION, MATERIAL FOR PREPARATION AND METHOD FOR THE PRODUCTION THEREOF |
WO2020206349A1 (en) * | 2019-04-05 | 2020-10-08 | Qidni Labs Inc. | Sorbent for use in renal therapy |
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US5681572A (en) * | 1991-10-18 | 1997-10-28 | Seare, Jr.; William J. | Porous material product and process |
US5686289A (en) * | 1993-10-08 | 1997-11-11 | The University Of Michigan | Method and compositions of a bioartificial kidney suitable for use in vivo or ex vivo |
WO2001024842A2 (en) * | 1999-10-05 | 2001-04-12 | Transkaryotic Therapies, Inc. | Hybrid matrices and hybrid matrix mixtures |
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US6419920B1 (en) * | 1995-10-25 | 2002-07-16 | Trans Karyotic Therapies, Inc. | Hybrid matrix implants and explants |
ATE260076T1 (de) * | 1996-09-05 | 2004-03-15 | Childrens Medical Center | Nierenprothese |
US6277574B1 (en) * | 1999-04-09 | 2001-08-21 | Incyte Genomics, Inc. | Genes associated with diseases of the kidney |
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2002
- 2002-05-17 JP JP2002588876A patent/JP4181877B2/ja not_active Expired - Fee Related
- 2002-05-17 US US10/478,170 patent/US20040235161A1/en not_active Abandoned
- 2002-05-17 WO PCT/JP2002/004778 patent/WO2002091955A1/ja active Application Filing
- 2002-05-17 CA CA2445785A patent/CA2445785C/en not_active Expired - Fee Related
- 2002-05-17 TW TW091110566A patent/TWI221421B/zh not_active IP Right Cessation
- 2002-05-17 KR KR1020037014699A patent/KR100864648B1/ko active IP Right Grant
- 2002-05-17 EP EP02769619A patent/EP1388327B1/en not_active Expired - Lifetime
- 2002-05-17 CN CNB028100514A patent/CN100335015C/zh not_active Expired - Fee Related
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- 2004-09-07 HK HK04106757A patent/HK1064024A1/xx not_active IP Right Cessation
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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US5681572A (en) * | 1991-10-18 | 1997-10-28 | Seare, Jr.; William J. | Porous material product and process |
US5686289A (en) * | 1993-10-08 | 1997-11-11 | The University Of Michigan | Method and compositions of a bioartificial kidney suitable for use in vivo or ex vivo |
WO2001024842A2 (en) * | 1999-10-05 | 2001-04-12 | Transkaryotic Therapies, Inc. | Hybrid matrices and hybrid matrix mixtures |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005094912A1 (ja) * | 2004-03-31 | 2005-10-13 | Cardio Incorporated | 血管新生作用が強化された移植片 |
JP2006042795A (ja) * | 2004-06-30 | 2006-02-16 | Hokkaido Univ | 細胞培養用基材、その製造方法及び細胞培養方法 |
JP4752047B2 (ja) * | 2004-06-30 | 2011-08-17 | 国立大学法人北海道大学 | 細胞培養用基材の製造方法及び細胞培養方法 |
WO2007029677A1 (ja) * | 2005-09-09 | 2007-03-15 | Gunze Limited | 組織再生用基材 |
US8889171B2 (en) | 2005-09-09 | 2014-11-18 | Gunze Limited | Tissue regeneration substrate |
WO2007119563A1 (ja) | 2006-03-28 | 2007-10-25 | Niigata University | ヒトメガリンの測定方法 |
Also Published As
Publication number | Publication date |
---|---|
KR20040004621A (ko) | 2004-01-13 |
TWI221421B (en) | 2004-10-01 |
EP1388327A1 (en) | 2004-02-11 |
CA2445785A1 (en) | 2002-11-21 |
US20040235161A1 (en) | 2004-11-25 |
JPWO2002091955A1 (ja) | 2004-08-26 |
CA2445785C (en) | 2012-03-20 |
HK1064024A1 (en) | 2005-01-21 |
CN100335015C (zh) | 2007-09-05 |
CN1509155A (zh) | 2004-06-30 |
KR100864648B1 (ko) | 2008-10-23 |
JP4181877B2 (ja) | 2008-11-19 |
EP1388327A4 (en) | 2009-08-19 |
EP1388327B1 (en) | 2012-02-22 |
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