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KR970705951A - Three-Dimensional Human Cell Culture in Heart Valve Structure and Its Use (THREE-DIMENSIONAL HUMAN CELL CULTURES ON CARDIAC VALVE FRANEWORKS AND THEIR USES) - Google Patents

Three-Dimensional Human Cell Culture in Heart Valve Structure and Its Use (THREE-DIMENSIONAL HUMAN CELL CULTURES ON CARDIAC VALVE FRANEWORKS AND THEIR USES) Download PDF

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KR970705951A
KR970705951A KR1019970701618A KR19970701618A KR970705951A KR 970705951 A KR970705951 A KR 970705951A KR 1019970701618 A KR1019970701618 A KR 1019970701618A KR 19970701618 A KR19970701618 A KR 19970701618A KR 970705951 A KR970705951 A KR 970705951A
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heart valve
stromal
stromal cells
cells
culture medium
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가일 케이. 노우톤
브리안 에이. 노우톤
안토니 에프. 퓨리치오
리. 케이. 란덴
죠안 질팅거
토드 디. 캠벨
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가일 케이. 노우톤
어드밴스드 티슈 사이언스, 인코퍼레이티드
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Publication of KR970705951A publication Critical patent/KR970705951A/en
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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
    • A61L27/3804—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
    • 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
    • A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
    • A61F2/06—Blood vessels
    • A61F2/062—Apparatus for the production of blood vessels made from natural tissue or with layers of living cells
    • 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
    • A61F2/24—Heart valves ; Vascular valves, e.g. venous valves; Heart implants, e.g. passive devices for improving the function of the native valve or the heart muscle; Transmyocardial revascularisation [TMR] devices; Valves implantable in the body
    • A61F2/2412—Heart valves ; Vascular valves, e.g. venous valves; Heart implants, e.g. passive devices for improving the function of the native valve or the heart muscle; Transmyocardial revascularisation [TMR] devices; Valves implantable in the body with soft flexible valve members, e.g. tissue valves shaped like natural valves
    • A61F2/2415—Manufacturing methods
    • 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
    • A61L27/3604—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 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3625—Vascular tissue, e.g. heart valves
    • 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
    • A61L27/3641—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 characterised by the site of application in the body
    • A61L27/3645—Connective tissue
    • 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
    • A61L27/3683—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 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • 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
    • A61L27/3843—Connective tissue
    • 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
    • A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels

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  • Health & Medical Sciences (AREA)
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  • Zoology (AREA)
  • Cardiology (AREA)
  • Urology & Nephrology (AREA)
  • Heart & Thoracic Surgery (AREA)
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  • Cell Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Pulmonology (AREA)
  • Manufacturing & Machinery (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Materials For Medical Uses (AREA)
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Abstract

본 발명은 배양배지에서 사람 섬유아세포를 이용하여 시험관에서 3차 배양물에서 상이한 세포와 조직을 생장시키는 방법에 관계한다. 본 발명에 따르면, 사람피부와 심장 섬유아세포를 포함하는 기질세포는 3차 구조 또는 뼈대에 접종시키고 생장시킨다. 사람 섬유아세포는 기질세포에 의해 연결된 세포간 공간을 가지는 3차 구조면에서 형성된 세포구조 해체된 심장벨브 또는 대동맥 벽과 소엽으로 구성된 기존의 돼지 매트릭스에 보충하게 대체되는 사람 매트릭스 단백질을 분비한다.The present invention relates to a method for growing different cells and tissues in a tertiary culture in a test tube using human fibroblasts in a culture medium. According to the present invention, stromal cells comprising human skin and cardiac fibroblasts are inoculated and grown in tertiary structure or skeleton. Human fibroblasts secrete human matrix proteins that are replenished to existing pig matrices composed of disassembled heart valves or aortic wall and lobules formed in a tertiary structure with intercellular space connected by stromal cells.

형성된 살아있는 기질조직은 생체내에 이식된 배양물에서 장시간 생존능과 세포증식을 유지하는데 필요한 서포트와 성장인자와 조절인자를 제공한다. 3차 배양배지에서 생장하였을 때 증식세포는 성숙하고 생체내 동일부분과 유사한 조직 성분을 형성하기 위해 분류된다.The formed living matrix tissue provides support and growth factors and regulatory factors necessary to maintain long-term viability and cell proliferation in vivo implanted cultures. When grown on a tertiary culture medium, proliferating cells are classified to form maturation and tissue components similar to the same in vivo.

Description

심장 판막 구조에서 3차원 사람 세포 배양물과 이의 용도(THREE-DIMENSIONAL HUMAN CELL CULTURES ON CARDIAC VALVE FRANEWORKS AND THEIR USES)Three-Dimensional Human Cell Culture in Heart Valve Structure and Its Use (THREE-DIMENSIONAL HUMAN CELL CULTURES ON CARDIAC VALVE FRANEWORKS AND THEIR USES)

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is a trivial issue, I did not include the contents of the text.

제1도는 돼지 동맥 소엽과 벽에 사람 피부 섬유아세포에 접종하여 합성된 자가방사된 단백질의 사진이다. 제2도는 a) (새로운 돼지 소엽 조직에 고유적인 심장 섬유아세포 핵은 색이 보라색으로 나타남); b) 계면활성제 그리고/또는 효소 추출한 돼지 소엽(화학 처리후에 돼지 세포핵이 감지되지 않음); c) 18주간 사람 섬유아세포로 배양된 계면 활성제 그리고/또는 효소 추출한 돼지 소엽(돼지 기질에 사람 피부 섬유아세포가 나타남)(헤마 토실린/에오신)(10x); 헤마토실린/에오신 착색된 조직부분의 사진이다, 제3도는 사람 피부 섬유아세포로 접종하고 4주간 배양시킨 돼지 혈소판의 사진이다, (헤마 토실린/에오신으로 착색) 제4도는 섬유아세포 유무하에 계면활성제 그리고/또는 효소추출한 소염의 세개 샘플 세트에서의 막대 그래프를 나타내는데 이때3H-티미닌이 결합된 섬유아세포로 성장된 소엽에서만 섬유아세포가 증식됨을 나타낸다, 제5도는 단백질 밴드를 나타내는 SPS 겔-지 가방사 그래프.Figure 1 is a photograph of self-radiated protein synthesized by inoculating human dermal fibroblasts into porcine arterial lobules and walls. FIG. 2 a) (cardiac fibroblast nuclei unique to new pig lobular tissue appear purple in color); b) pork lobes extracted with surfactants and / or enzymes (no pig cell nuclei detected after chemical treatment); c) Surfactants cultured with human fibroblasts for 18 weeks and / or enzyme-extracted porcine lobes (human skin fibroblasts appear in porcine matrix) (hematocillin / eosin) (10x); 3 is a photograph of a pig platelet inoculated with human dermal fibroblast and cultured for 4 weeks (coloring with hematocillin / eosin). Fig. 4 is a photograph of a hematocillin / eosin- Figure 3 shows a bar graph in three sample sets of active agent and / or enzyme-extracted anti-inflammatory, wherein fibroblasts are proliferated only in lobules grown with 3 H-thyminin-conjugated fibroblasts. Figure 5 shows SPS gel- The graph of the paper bag.

Claims (22)

시험관에서 준비된 살아있는 기질 세포-콜로니화된 심장 판막에 있어서 심장 판막에 접종된 기질 세포에 의해 자연적으로 분비되는 기질세포와 연결조직 단백질로 구성되어 심장판막은 기질 세포에 의해 연결된 세포간 공간을 가지는 3차 구조를 형성하기 위한 살아있는 세포로 재구성되는 것을 특징으로 하는 심장판막.Living stromal cells prepared in vitro - composed of stromal cells and connective tissue proteins that are naturally secreted by stromal cells in the heart valve in the colonized heart valve, and heart valves have intercellular space connected by stromal cells 3 A heart valve characterized by being reconstituted into living cells for forming a car structure. 제1항에 있어서, 기질세포는 섬유아세포인 것을 특징으로 하는 심장판막.The heart valve according to claim 1, wherein the stromal cells are fibroblasts. 제1항에 있어서, 기질세포는 사람 피부 섬유아세포인 것을 특징으로 하는 심장판막.The heart valve according to claim 1, wherein the stromal cell is human skin fibroblast. 제1항에 있어서, 기질세포는 사람 심장 섬유아세포인 것을 특징으로 하는 심장판막.The heart valve according to claim 1, wherein the stromal cells are human cardiac fibroblasts. 제1항에 있어서, 심장판막은 돼지의 것인 것을 특징으로 하는 심장판막.The heart valve according to claim 1, wherein the heart valve is of a pig. 제5항에 있어서, 심장판막은 기질세포 접종전에 세포구조 해체되는 것을 특징으로 하는 심장판막.6. The heart valve according to claim 5, wherein the heart valve is disassembled before the stromal cell inoculation. 제6항에 있어서, 심장판막은 효소와 계면활성제 처리에 의해 세포 구조해체되거나 냉동/해동에 의해 살아있지 않게 만드는 것을 특징으로 하는 심장판막.The heart valve according to claim 6, wherein the heart valve is disrupted by the disintegration of the cell structure or freezing / thawing by treatment with an enzyme and a surfactant. 제1항에 있어서, 심장판막은 생체분해 가능한 물질로 구성된 것을 특징으로 하는 심장판막.The heart valve according to claim 1, wherein the heart valve is made of a biodegradable material. 제8항에 있어서, 생체분해 가능한 물질은 폴리글리콜산, 고양이 내장 봉합사, 콜라겐, 셀룰로오즈, 겔라틴, 히알루론산 또는 폴리하이드록시알카노네이트로 구성된 것을 특징으로 하는 심장판막.9. The heart valve of claim 8, wherein the biodegradable material comprises polyglycolic acid, a built-in suture of a cat, collagen, cellulose, gelatin, hyaluronic acid or polyhydroxyalkanoate. 시험관에서 준비된 살아있는 기질 세포-피복된 심장판막을 만드는 방법에 있어서, 배양배지에서 심장판막에 접종된 배양된 기질세포를 구성되고 3차 구조를 형성하기 위한 심장판막에 부착된 기질세포에 의해 기질세포와 연결조직 사람 단백질은 자연적으로 분비되는 것을 특징으로 하는 방법.A method for producing a living stromal cell-coated heart valve prepared in vitro, comprising culturing the stromal stem cells inoculated in the heart valve in a culture medium and culturing the stromal cells And the connective tissue protein is secreted naturally. 제10항에 있어서, 기질세포는 섬유아세포인 것을 특징으로 하는 방법.11. The method of claim 10, wherein the stromal cell is a fibroblast. 제10항에 있어서, 기질세포는 사람 피부 섬유아세포인 것을 특징으로 하는 심장판막.11. The heart valve of claim 10, wherein the stromal cells are human dermal fibroblasts. 제10항에 있어서, 기질세포는 사람 심장 섬유아세포인 것을 특징으로 하는 방법.11. The method of claim 10, wherein the stromal cells are human cardiac fibroblasts. 제10항에 있어서, 심장판막은 돼지의 것인 것을 특징으로 하는 심장판막.11. The heart valve according to claim 10, wherein the heart valve is of a pig. 제14항에 있어서, 심장판막은 기질세포 접종전에 세포구조 해체되는 것을 특징으로 하는 방법.15. The method of claim 14, wherein the heart valve is disassembled prior to stromal cell inoculation. 제15항에 있어서, 심장판막은 효소와 계면활성제 처리에 의해 세포 구조해체되거나 냉동/해동에 의해 살아있지 않게 만드는 것을 특징으로 하는 방법.16. The method of claim 15, wherein the heart valve is disrupted by enzymatic and surfactant treatment or by freezing / thawing. 제10항에 있어서, 심장판막은 생체분해 가능한 물질로 구성된 것을 특징으로 하는 방법.11. The method of claim 10, wherein the heart valve is comprised of a biodegradable material. 제17항에 있어서, 생체분해 가능한 물질은 폴리글리콜산, 고양이 내장 봉합사, 콜라겐, 셀룰로오즈, 겔라틴, 히알루론산 또는 폴리하이드록시 알카노네이트로 구성된 것을 특징으로 하는 방법.18. The method of claim 17, wherein the biodegradable material comprises polyglycolic acid, a cat internal suture, collagen, cellulose, gelatin, hyaluronic acid or polyhydroxyalkanoate. 제10항에 있어서, 배양배지는 정적상태에서 유지되는 것을 특징으로 하는 방법.11. The method of claim 10, wherein the culture medium is maintained in a static state. 제10항에 있어서, 배양배지는 순환시키는 것으로 동적 상태에서 유지되는 것을 특징으로 하는 방법.11. The method of claim 10, wherein the culture medium is maintained in a dynamic state by circulation. 제10항에 있어서, 배양배지의 성장인자, 자연 또는 변형된 혈액 생성물 또는 생체 활성분자의 이용으로 구성된 것을 특징으로 하는 방법.11. The method of claim 10, comprising the use of growth factors, natural or modified blood products or bioactive molecules of the culture medium. 살아있는 기질세포 콜로니 형성된 심장판막의 이식방법에 있어서; a) 세포구조 해체된 심장판막에 기질 세포를 접종하고; b) 기질세포를 배양하여 시험관에서 증식시키고; 그리고 c) 생체에서 기질세포-피복된 사람화된 심장판막을 이식하는 것으로 구성된 것을 특징으로 하는 방법.A method for transplanting a living stromal cell colony formed heart valve; a) inoculating stromal cells to a heart valve with disrupted cell structure; b) incubation of stromal cells and propagation in vitro; And c) implanting stromal cell-coated humanized heart valves in vivo. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: It is disclosed by the contents of the first application.
KR1019970701618A 1994-09-12 1995-09-08 Three-Dimensional Human Cell Culture in Heart Valve Structure and Its Use (THREE-DIMENSIONAL HUMAN CELL CULTURES ON CARDIAC VALVE FRANEWORKS AND THEIR USES) Ceased KR970705951A (en)

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