CN114617671A - 一种光控心肌器官膜片的制备方法及应用 - Google Patents
一种光控心肌器官膜片的制备方法及应用 Download PDFInfo
- Publication number
- CN114617671A CN114617671A CN202011460631.3A CN202011460631A CN114617671A CN 114617671 A CN114617671 A CN 114617671A CN 202011460631 A CN202011460631 A CN 202011460631A CN 114617671 A CN114617671 A CN 114617671A
- Authority
- CN
- China
- Prior art keywords
- myocardial
- light
- film
- organ
- operated
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 230000002107 myocardial effect Effects 0.000 title claims abstract description 64
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000000463 material Substances 0.000 claims abstract description 32
- 239000012528 membrane Substances 0.000 claims abstract description 15
- 210000000056 organ Anatomy 0.000 claims abstract description 13
- 210000001519 tissue Anatomy 0.000 claims description 13
- 239000011521 glass Substances 0.000 claims description 12
- 210000004027 cell Anatomy 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 9
- 102000008186 Collagen Human genes 0.000 claims description 8
- 108010035532 Collagen Proteins 0.000 claims description 8
- 229920001436 collagen Polymers 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 238000007605 air drying Methods 0.000 claims description 4
- DMLAVOWQYNRWNQ-UHFFFAOYSA-N azobenzene Chemical compound C1=CC=CC=C1N=NC1=CC=CC=C1 DMLAVOWQYNRWNQ-UHFFFAOYSA-N 0.000 claims description 4
- 229940114081 cinnamate Drugs 0.000 claims description 4
- 239000004973 liquid crystal related substance Substances 0.000 claims description 4
- WBYWAXJHAXSJNI-VOTSOKGWSA-M trans-cinnamate Chemical compound [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 claims description 4
- AAAQKTZKLRYKHR-UHFFFAOYSA-N triphenylmethane Chemical compound C1=CC=CC=C1C(C=1C=CC=CC=1)C1=CC=CC=C1 AAAQKTZKLRYKHR-UHFFFAOYSA-N 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 239000008280 blood Substances 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- 238000005520 cutting process Methods 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 235000019441 ethanol Nutrition 0.000 claims description 3
- 238000011156 evaluation Methods 0.000 claims description 3
- 210000002744 extracellular matrix Anatomy 0.000 claims description 3
- 239000003517 fume Substances 0.000 claims description 3
- 239000003292 glue Substances 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 238000002294 plasma sputter deposition Methods 0.000 claims description 3
- 238000003892 spreading Methods 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 231100000041 toxicology testing Toxicity 0.000 claims description 3
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 claims description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 claims description 2
- -1 benzspiropyran Chemical compound 0.000 claims description 2
- 230000002349 favourable effect Effects 0.000 claims description 2
- 238000000034 method Methods 0.000 claims description 2
- 229920000642 polymer Polymers 0.000 claims description 2
- 238000003255 drug test Methods 0.000 abstract description 6
- 239000008204 material by function Substances 0.000 abstract description 4
- 231100000820 toxicity test Toxicity 0.000 abstract description 4
- 230000002269 spontaneous effect Effects 0.000 abstract description 3
- 238000005286 illumination Methods 0.000 abstract description 2
- 230000009191 jumping Effects 0.000 abstract description 2
- 238000012986 modification Methods 0.000 abstract description 2
- 230000004048 modification Effects 0.000 abstract description 2
- 210000004165 myocardium Anatomy 0.000 description 6
- 210000004413 cardiac myocyte Anatomy 0.000 description 5
- 238000010009 beating Methods 0.000 description 4
- 239000004205 dimethyl polysiloxane Substances 0.000 description 4
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 4
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 4
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 4
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 4
- 239000012620 biological material Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- KYNSBQPICQTCGU-UHFFFAOYSA-N Benzopyrane Chemical compound C1=CC=C2C=CCOC2=C1 KYNSBQPICQTCGU-UHFFFAOYSA-N 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000007385 chemical modification Methods 0.000 description 2
- 230000033001 locomotion Effects 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000000887 hydrating effect Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000010349 pulsation Effects 0.000 description 1
- 210000001567 regular cardiac muscle cell of ventricle Anatomy 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- 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
- A61F2/08—Muscles; Tendons; Ligaments
-
- 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
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0657—Cardiomyocytes; Heart cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5061—Muscle 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
- A61F2/08—Muscles; Tendons; Ligaments
- A61F2002/0894—Muscles
-
- 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
- A61L2430/20—Materials or treatment for tissue regeneration for reconstruction of the heart, e.g. heart valves
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2529/00—Culture process characterised by the use of electromagnetic stimulation
- C12N2529/10—Stimulation by light
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/54—Collagen; Gelatin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/10—Screening for compounds of potential therapeutic value involving cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Transplantation (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Cardiology (AREA)
- Pathology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Vascular Medicine (AREA)
- Organic Chemistry (AREA)
- Heart & Thoracic Surgery (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Rheumatology (AREA)
- Food Science & Technology (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- General Engineering & Computer Science (AREA)
- Orthopedic Medicine & Surgery (AREA)
Abstract
本发明属于生物医学工程与功能材料、器官芯片技术领域，具体涉及一种光控心肌器官膜片的制备方法及生物应用。本发明利用光响应材料制备超薄膜片，经过表面修饰，促进心肌细胞粘附和生长，制备的心肌器官膜片具有随着心肌细胞自发跳动的功能；同时也可以光照刺激膜片运动，被动牵拉心肌细胞。所述光控心肌器官膜片可以实现心肌组织工程，心肌药物测试以及心肌毒性测试等领域。
Description
技术领域
本发明属于生物医学工程与功能材料、器官芯片技术领域，涉及一种光控心肌器官膜片的制备方法及生物应用。本发明的光控心肌器官膜片适用于实现心肌组织工程，心肌药物测试以及心肌毒性测试等领域。
背景技术
现有技术公开了生物材料和功能材料在心肌组织工程，心肌器官芯片构建等方面发挥重要的作用。现有的生物材料和功能材料主要包括胶原，明胶，海藻酸钠等细胞外基质和生物高分子等，研究显示，这些生物材料可以作为心肌细胞的支撑，心肌细胞可以带动这些材料跳动，制备的工程化的心肌组织可以用于进行心肌修补，心肌药物测试以及毒性分析等。
光响应材料是含有偶氮苯、苯并螺吡喃、三苯基甲烷、肉桂酸酯的液晶材料，是一种新型的可以制备各种不同功能的微流控领域的材料，其特征在于可以实现光控材料发生形变，在密闭的微管中推动液体流动等特性，但该材料在心肌组织工程等领域的作用尚未开发。
为了拓展该材料在生物医药技术领域的应用，本发明利用所述材料光控致形变的特性，制备一种光控心肌器官膜片，制备的心肌组织薄膜可以在光控条件下被动形变，可以用于研究心肌细胞被动牵拉引起的病理改变，同时也可以实现心肌自发跳动引起的薄膜跳动，用于心肌组织工程，药物测试，毒性测试等领域。
发明内容
本发明的目的在于，基于现有技术的现状，提供一种光控心肌器官膜片。本发明利用新的光致形变材料的薄膜制备的新方法，采用有关光控致形变特性的材料，制备光控心肌器官膜片，制备的心肌组织薄膜可以在光控条件下被动形变，可以用于研究心肌细胞被动牵拉引起的病理改变，同时也可以实现心肌自发跳动引起的薄膜跳动，用于心肌组织工程，药物测试，毒性测试等领域，并在此基础上实现心肌组织工程。
本发明提供了一种光控心肌器官膜片的制备方法，其特征在于：所述光控心肌器官膜片的材料为光响应高分子聚合物形成的超薄半透明膜经过表面修饰，可以实现心肌细胞的粘附和生长；该膜片可以随着心肌的跳动而运动，也可以实现光照对心肌膜片运动的控制。
本发明所述的光响应材料为具有光致形变能力的含有偶氮苯、苯并螺吡喃、三苯基甲烷、肉桂酸酯的液晶材料；所述光敏感材料不限于上述几种，包括在此基础上进行的化学修饰等新材料。
本发明所述的控材料不限于含有偶氮苯、苯并螺吡喃、三苯基甲烷、肉桂酸酯的液晶材料，也包括其他在此基础上进行的其他化学修饰后产生的光敏感材料。
本发明所述的光响应薄膜通过下述过程制备：
(1)将合成的光响应材料用甲苯溶解完全，浓度范围在1-10％；
(2)将一定体积的液态的光响应材料滴加在干净的玻璃片上，以血涂片的方式用另外一个玻璃片将液滴涂开，通风橱内风干；
(3)根据制备膜厚度的不同要求，可以重复(2)步骤，制备不同厚度和透明度的光控薄膜
(4)将载有薄膜的玻璃片放在无水乙醇中，轻轻掀起周边，薄膜脱落，在无水乙醇中铺展开；
(5)用新的载玻片轻轻捞起薄膜，常温风干；
(6)氧等离子溅射5分钟亲水处理后，加入胶原或其他细胞外基质胶修饰表面；所述胶原浓度范围在30-80ug/ml；也可以使用细胞外基质胶原；
(7)用锐利的刀片将将薄膜切成需要的大小；固定两侧；
(8)75％酒精灭菌30分钟；备用。
本发明的光控心肌器官膜片可用于心肌组织工程，其制备方法为，接种心肌细胞前，利用PBS将薄膜水化，待细胞贴壁，跳动以后，去除薄膜一侧固定，使得一侧呈游离状态，利于薄膜运动。
所述薄膜表面修饰的胶原蛋白浓度范围在30-80ug/ml。
本发明的光控心肌器官薄膜可制备用于心肌组织工程，心肌毒性测试，心肌药物评价制品，应用于心肌组织工程，心肌毒性测试，心肌药物评价等领域。
附图说明
图1.光控薄膜制备流程图。
图2.制备的光控心肌器官膜片，
其中，A图中的1和2为固定膜的PDMS条，通过双面胶或PDMS胶水固定；3为光敏感材料制备的薄膜；4为小鼠心肌细胞；B.心肌细胞贴壁生长48小时候，去除2固定物；薄膜随着心肌细胞的收缩而发生卷曲运动。
具体实施方法
实施例1制备光控薄膜
(1)将合成的光响应材料用甲苯溶解完全；浓度范围为2％
(2)将30微升的液态的光响应材料滴加在干净的玻璃片上，以血涂片的方式用另外一个玻璃片将液滴涂开，通风橱内风干；
(3)根据制备膜厚度的不同要求，重复(2)步骤3次，制备光控薄膜厚度在100微米左右。
(4)将载有薄膜的玻璃片放在无水乙醇中，轻轻掀起周边，薄膜脱落，在无水乙醇中铺展开；
(5)用新的载玻片轻轻捞起薄膜，常温风干；
(6)氧等离子溅射5分钟亲水处理后，加入40ug/ml胶原容易修饰表面60分钟，室温风干；
(7)用锐利的刀片将将薄膜切成2毫米宽度，5毫米长度的形状；用PDMS固定两侧；
(8)75％酒精灭菌30分钟；放在培养皿内备用。
实施例2制备光控心肌器官膜片
(1)提取新生乳鼠的心室肌细胞，培养在培养皿中；
(2)将制备的光控薄膜用PBS清洗一次，加入心肌细胞培养基，高糖DMEM含有10％FBS和青链霉素各100单位/毫升；
(3)将心肌细胞消化成单细胞悬液，轻轻的滴加在薄膜上，静止培养2-6小时，促进心肌细胞贴壁；
(4)待心肌细胞贴壁后，补加新鲜培养基，继续培养24-48小时；
(5)心肌细胞跳动后，去除一侧固定的PDMS，将薄膜条分离开，一段游离，促进心肌膜的跳动；
(6)可以对跳动的心肌膜进行光控操作，可以对心肌膜进行药物测试。
Claims (7)
1.一种光控心肌器官膜片，其特征在于，通过下述方法制备：
(1)将合成的光响应材料用甲苯溶解完全，浓度范围在1-10％；
(2)将一定体积的液态的光响应材料滴加在干净的玻璃片上，以血涂片的方式用另外一个玻璃片将液滴涂开，通风橱内风干；
(3)根据制备膜厚度的不同要求，重复(2)步骤，制备不同厚度和透明度的光控薄膜；
(4)将载有薄膜的玻璃片置无水乙醇中，掀起周边，薄膜脱落，在无水乙醇中铺展开；
(5)用载玻片捞起薄膜，常温风干；
(6)氧等离子溅射5分钟亲水处理后，加入胶原或细胞外基质胶修饰表面；
(7)用锐利刀片将薄膜切成需要的大小；固定两侧；
(8)75％酒精灭菌30分钟；备用。
2.按权利要求1所述的光控心肌器官膜片，其特征在于，所述的光响应材料为光响应高分子聚合物形成的超薄半透明膜经过表面修饰的具光控致形变特性的材料。
3.按权利要求1或2所述的光控心肌器官膜片，其特征在于，所述的光响应材料为具有光致形变能力的含有偶氮苯、苯并螺吡喃、三苯基甲烷或肉桂酸酯的液晶材料。
4.按权利要求1或2所述的光控心肌器官膜片，其特征在于，所述的光控材料还包括其他化学修饰后产生的光敏感材料。
5.权利要求1所述的光控心肌器官膜片在制备用于心肌组织工程制品中的用途；所述的心肌组织工程制品使用时，包括，接种心肌细胞前，利用PBS将所述薄膜水化，待细胞贴壁，跳动以后，去除薄膜一侧固定，使得一侧呈游离状态，利于薄膜运动。
6.按权利要求5所述的用途，其特征在于，所述薄膜表面修饰胶原蛋白，其浓度范围在30-80ug/ml。
7.权利要求1所述的光控心肌器官膜片在制备用于心肌毒性测试，心肌药物评价制品中的用途。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011460631.3A CN114617671B (zh) | 2020-12-12 | 2020-12-12 | 一种光控心肌器官膜片的制备方法及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011460631.3A CN114617671B (zh) | 2020-12-12 | 2020-12-12 | 一种光控心肌器官膜片的制备方法及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114617671A true CN114617671A (zh) | 2022-06-14 |
| CN114617671B CN114617671B (zh) | 2024-06-18 |
Family
ID=81894781
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011460631.3A Active CN114617671B (zh) | 2020-12-12 | 2020-12-12 | 一种光控心肌器官膜片的制备方法及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114617671B (zh) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02211865A (ja) * | 1989-02-10 | 1990-08-23 | Kao Corp | 細胞培養支持体材料 |
| JP2003033177A (ja) * | 2001-07-24 | 2003-02-04 | Mitsuo Okano | 高密度細胞アレイ用基板、製造法、及びその利用方法 |
| CN104931683A (zh) * | 2015-05-21 | 2015-09-23 | 清华大学 | 一种心肌组织传感器及心肌组织芯片的制备方法 |
| US20160067284A1 (en) * | 2013-05-17 | 2016-03-10 | Terumo Kabushiki Kaisha | Method for producing sheet-shaped cell culture |
| US20170247658A1 (en) * | 2014-11-12 | 2017-08-31 | Terumo Kabushiki Kaisha | Myocardial cell sheet |
-
2020
- 2020-12-12 CN CN202011460631.3A patent/CN114617671B/zh active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02211865A (ja) * | 1989-02-10 | 1990-08-23 | Kao Corp | 細胞培養支持体材料 |
| JP2003033177A (ja) * | 2001-07-24 | 2003-02-04 | Mitsuo Okano | 高密度細胞アレイ用基板、製造法、及びその利用方法 |
| US20160067284A1 (en) * | 2013-05-17 | 2016-03-10 | Terumo Kabushiki Kaisha | Method for producing sheet-shaped cell culture |
| US20170247658A1 (en) * | 2014-11-12 | 2017-08-31 | Terumo Kabushiki Kaisha | Myocardial cell sheet |
| CN104931683A (zh) * | 2015-05-21 | 2015-09-23 | 清华大学 | 一种心肌组织传感器及心肌组织芯片的制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114617671B (zh) | 2024-06-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bakirci et al. | Cell sheet based bioink for 3D bioprinting applications | |
| Rayatpisheh et al. | Combining cell sheet technology and electrospun scaffolding for engineered tubular, aligned, and contractile blood vessels | |
| Cooperstein et al. | Biological cell detachment from poly (N-isopropyl acrylamide) and its applications | |
| US11371021B2 (en) | Encapsulation and cardiac differentiation of hiPSCs in 3D PEG-fibrinogen hydrogels | |
| EP2781594A1 (en) | Cell culture substrate, and method for manufacturing same | |
| JP2008263863A (ja) | 細胞培養支持体とその製造方法 | |
| JP5396803B2 (ja) | 細胞培養基材及び細胞培養方法 | |
| CN107899086A (zh) | 一种透明质酸寡糖修饰的胶原蛋白纳米纤维血管修复材料及其制备方法 | |
| Gugerell et al. | Adipose-derived stem cells cultivated on electrospun l-lactide/glycolide copolymer fleece and gelatin hydrogels under flow conditions–aiming physiological reality in hypodermis tissue engineering | |
| Kim et al. | Recent advances in engineered stem cell-derived cell sheets for tissue regeneration | |
| CN106924817B (zh) | 一种超薄载体细胞片及其制备方法 | |
| CN105566567A (zh) | 硬度可调节的温敏聚合物材料及其制备方法和应用 | |
| Elomaa et al. | In vitro vascularization of hydrogel-based tissue constructs via a combined approach of cell sheet engineering and dynamic perfusion cell culture | |
| Yong et al. | Interdisciplinary approaches to advanced cardiovascular tissue engineering: ECM-based biomaterials, 3D bioprinting, and its assessment | |
| CN104225670A (zh) | 可控性疏水细菌纤维素-玉米醇溶蛋白复合膜的制备方法 | |
| CN108187142A (zh) | 一种干细胞在生物组织修复中的应用 | |
| Akiyama et al. | Temperature-responsive polymers for cell culture and tissue engineering applications | |
| Seo et al. | Recent and future strategies of mechanotherapy for tissue regenerative rehabilitation | |
| Marei et al. | Assessment of parylene C thin films for heart valve tissue engineering | |
| CN103898055A (zh) | 不同基底硬度体外细胞培养平台的建立方法 | |
| CN107805625A (zh) | 一种石墨烯培养基底及其制备方法和在多能干细胞培育、分化及移植方面的应用 | |
| CN114617671A (zh) | 一种光控心肌器官膜片的制备方法及应用 | |
| CN103877613B (zh) | 基于微冰胶构建注射型三维细胞微环境的系统和方法 | |
| CN110746548B (zh) | 一种制备细胞支架的水凝胶 | |
| WO2018116902A1 (ja) | 細胞培養基材 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant |