CN108939157A - The preparation method of natural tubulose small intestine acellular matrix film - Google Patents
The preparation method of natural tubulose small intestine acellular matrix film Download PDFInfo
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- CN108939157A CN108939157A CN201810839013.6A CN201810839013A CN108939157A CN 108939157 A CN108939157 A CN 108939157A CN 201810839013 A CN201810839013 A CN 201810839013A CN 108939157 A CN108939157 A CN 108939157A
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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
- A61L27/3604—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
- A61L27/3633—Extracellular matrix [ECM]
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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
- A61L27/3641—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
- A61L27/3679—Hollow organs, e.g. bladder, esophagus, urether, uterus, intestine
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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
- A61L27/3683—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
- A61L27/3691—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 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
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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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/22—Materials or treatment for tissue regeneration for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
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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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
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Abstract
The present invention provides a kind of preparation method of natural tubulose small intestine acellular matrix film, belongs to medical biomaterial technical field.This method comprises: the small intestine intercepted in animals is cleaned；Small intestine after cleaning up is placed in closed container, it is rapidly frozen in -80 DEG C of liquid nitrogen, it takes out after half an hour and is nested on annular tube shaped bracket after thawing, placenta percreta is destroyed with scissors, tunica serosa intestini tenuis layer is cut off, is struck off with tunica serosa intestini tenuis layer of the poly (methyl methacrylate) plate to destruction, the mucous layer, submucosa and muscle layer of small intestine are struck off after overturning small intestine, to small intestine, basal cell is removed, and obtains single-layer tubular small intestine acellular matrix film.Obtained single layer and multilayer tubular small intestine acellular matrix film are subjected to freeze-drying sealing, sterilizing preservation, obtain the natural tubulose small intestine acellular matrix film of different-thickness.Preparation method of the invention is simple, and obtained small intestine acellular matrix film is natural tubulose.
Description
Technical field
The invention belongs to medical biomaterial technical fields, and in particular to a kind of natural tubulose small intestine acellular matrix film system
Preparation Method.
Background technique
Small intestine acellular matrix film (ECM) biological sticking patch, refers to and is derived from of the same race or xenogenesis tissue, through de- cell processing
The three-dimensional for removing the various cells contained in tissue and completely retaining extracellular matrix (extracellular matr ix, ECM)
Frame structure and the material for being used to repair body tissue.When carrying out histoorgan reparation using ECM biological sticking patch or rebuilding,
ECM can play the role of " mold ".ECM biological sticking patch transmits biological information to the ECM receptor of host, passes through integrin, layer
Fibronectin receptor etc. induces and corresponding histoorgan cell is stimulated to adhere on its surface, migrates from edge to center, and lure
Guided cell is along the shape differentiation and growth of " mold ", and the final regeneration for realizing histoorgan, at the same time, ECM is gradually dropped
It solves and fully absorbs, the noresidue in host.
The clinical application range of ECM biological sticking patch is extensive, can be applied to abdominal hernia, pleura, skull, nerve, endomeninx,
The reparation of alimentary canal and skin etc..But the biological sticking patch as made of natural cell epimatrix material is excessively soft, machine
Tool performance is poor, brings many inconvenience for surgical procedure, in the reparation of Various Tissues, cell epimatrix material degradation difficult to realize
Speed is consistent with regeneration speed, often leads to graft failure because material degradation is too fast.In recent years, with science and technology it is fast
Speed development, for the improvement for how avoiding the generation proposition of the above problem many in the preparation of ECM biological sticking patch.Usually to material
Chemical crosslinking processing is carried out, the external research to the preparation of tubulose ECM biological sticking patch is more, when preparing tubulose ECM biological sticking patch,
Foreign countries mostly use sheet ECM biological sticking patch to be sewn into tubulose, or sheet ECM biological sticking patch is surrounded column as twining bandage
Shape mold is wound in tubulose.But this tubulose ECM biological sticking patch by sewing or being entwined, sutural margin it is reliable
The uniformity coefficient of property, the slickness of inside pipe wall and tube wall all can reparation to tissue or reconstruction impact.China is to ECM
The research and application of biological sticking patch are started late, and preparation process is relatively backward, especially for natural tubulose ECM biological sticking patch
Preparation and application study are still in blank.
Summary of the invention
The purpose of the invention is to provide a kind of natural tubulose small intestine acellular matrix membrane preparation method, the preparation method
Simply, the small intestine acellular matrix film obtained is natural tubulose, plays the role of good foundation to producing for artificial tubular organ.
Present invention firstly provides a kind of preparation methods of natural tubulose small intestine acellular matrix film, this method comprises:
Step 1: the small intestine intercepted in animals being immersed in physiological saline and is cleaned, small after cleaning up
Intestines are placed in closed container, are rapidly frozen in -80 DEG C of liquid nitrogen, and it is spare that defrosting is taken out after half an hour；
Step 2: the small intestine after defrosting is nested on annular tube shaped bracket, destroys placenta percreta with scissors, cuts off small intestine slurry
Then film layer is struck off with tunica serosa intestini tenuis layer of the poly (methyl methacrylate) plate to destruction, the mucous layer of small intestine is struck off after overturning small intestine, is glued
Film lower layer and muscle layer, to small intestine, basal cell is removed, and obtains single-layer tubular small intestine acellular matrix film；
Step 3: the multilayer tubular small intestine acellular matrix film that step 2 obtains is subjected to freeze-drying sealing, sterilizing preservation, is obtained
To the natural tubulose small intestine acellular matrix film of single layer.
Preferably, the animal of the step one is pig, ox, sheep, mouse, horse.
Preferably, the cleaning of the step one is cleaned in 4 DEG C of physiological saline.
Preferably, the step one refrigerating process specifically: the small intestine after cleaning up is placed in closed container,
It is rapidly frozen in -80 DEG C of liquid nitrogen, takes out and thaw after half an hour.
Preferably, the step three freeze temperature is -80 DEG C, and freeze-drying time is for 24 hours.
Preferably, the sterilizing of the step four is the multilayer tubular small intestine acellular matrix film that will be sealed through straight line
Accelerate with 8000 ladd radiological dose radiation sterilizations.
The present invention provides the natural tubulose small intestine acellular matrix film that above-mentioned preparation method obtains.
Beneficial effects of the present invention
Present invention firstly provides a kind of preparation method of natural tubulose small intestine acellular matrix film, method of the invention can be with
The small intestine acellular matrix film of natural tubulose is directly prepared.The small intestine acellular matrix film of the tubulose due to be it is natural,
Tubulose, it can be used as the bracket of cell growth.Natural tubulose ECM of the invention has the characteristics that tubulose uniformity is good, and ECM is without bright
It is aobvious to destroy, illustrate that the preparation method can produce good natural tubulose small intestine acellular matrix film.
The present invention also provides above-mentioned preparation methods to obtain natural tubulose small intestine acellular matrix film, and the natural tubulose small intestine is de-
Cellular matrix film, it is identical with sheet ECM other performance for local organization, except that natural tubulose is without suture, base
Plasma membrane is lateral, longitudinal, etc. all directions stretcher strain it is all identical as small intestine, and the ECM after suturing is made due to abutment joint credit union
It is deformed in different directions drawing process at local host film limited.Therefore natural tubulose ECM, for human artificial oesophagus and its
He plays into gate action at producing for artificial pipe, lays the foundation for the research of next step artificial esophagus.
Detailed description of the invention
Fig. 1 is the photograph before the natural tubulose chitterlings acellular matrix film inflation of single layer that the embodiment of the present invention 1 is prepared
Piece figure.
Fig. 2 is the photograph after the natural tubulose chitterlings acellular matrix film inflation of single layer that the embodiment of the present invention 1 is prepared
Piece figure.
Fig. 3 is after the natural tubulose chitterlings acellular matrix film of single layer that the embodiment of the present invention 1 is prepared is dried in vacuo
Photo figure.
Fig. 4 is that the natural tubulose chitterlings acellular matrix film for the single layer that the embodiment of the present invention 1 is prepared puts 1000 times
Scanned photograph under lower Electronic Speculum.
Fig. 5 is that the natural tubulose chitterlings acellular matrix film for the single layer that the embodiment of the present invention 1 is prepared puts 5000 times
Scanned photograph under lower Electronic Speculum.
Specific embodiment
Present invention firstly provides a kind of preparation methods of natural tubulose small intestine acellular matrix film, this method comprises:
Step 1: the small intestine intercepted in animals being immersed in physiological saline and is cleaned, small after cleaning up
Intestines are placed in closed container, are rapidly frozen in -80 DEG C of liquid nitrogen, and it is spare that defrosting is taken out after half an hour；
Step 2: the small intestine after defrosting is nested on annular tube shaped bracket, destroys placenta percreta with scissors, as much as possible
Tunica serosa intestini tenuis layer uniformly is cut off, is then struck off with tunica serosa intestini tenuis layer of the poly (methyl methacrylate) plate to destruction；After scraped clean, turn over
Turn the mucous layer, submucosa and muscle layer that strike off small intestine after small intestine again, basal cell removes to small intestine, and it is small to obtain single-layer tubular
Intestines acellular matrix film.
Step 3: the single-layer tubular small intestine acellular matrix film that step 2 obtains is subjected to freeze-drying sealing, sterilizing preservation, is obtained
To the natural tubulose small intestine acellular matrix film of single layer.
According to the present invention, described immerse the small intestine intercepted in animals in physiological saline is cleaned, preferably
Immerse in 4 DEG C of physiological saline and clean immediately after small intestine is taken out, the animal is not particularly limited, preferably pig, ox,
Sheep, mouse or horse.
According to the present invention, the small intestine that small intestine is immersed after physiological saline cleaning is rapidly frozen, preferably will be clear
Small intestine after wash clean is placed in closed container, is rapidly frozen in -80 DEG C of liquid nitrogen, cooling time is half an hour；
According to the present invention, the small intestine after defrosting is nested on annular tube shaped bracket, the bracket is not particularly limited,
It can play the role of supporting in small intestine, then cut off tunica serosa intestini tenuis layer with scissors, uniformly cut off osculum as much as possible,
Then it is struck off with tunica serosa intestini tenuis layer of the poly (methyl methacrylate) plate to destruction.The source of the poly (methyl methacrylate) plate be it is commercially available, it is organic
The edge notches of glass plate wants moderate, cannot be too sharp keen, otherwise has damage to matrix membrane, cannot excessively targe, cell strike off
It is incomplete., firmly can not be too quickly when being struck off with poly (methyl methacrylate) plate, matrix membrane can be damaged by overexerting, then be struck off with same method
Mucous layer, submucosa and the muscle layer of small intestine, to small intestine, basal cell is removed.When inorganization falls off, sample is sufficiently rinsed,
Row strikes off again, and leaves and takes scrapings, and the liquid of scraping, and the scrapings left and taken and the liquid of scraping are centrifuged, after taking centrifugation
Sediment smear, dyeing, under the microscope, such as cell-free ingredient strikes off completion to high power, obtains single-layer tubular small intestine and takes off cell base
Plasma membrane.
According to the present invention, multilayer tubular small intestine acellular matrix film obtained above is subjected to freeze-drying sealing, sterilizing preservation,
Obtain the natural tubulose small intestine acellular matrix film of single layer.The freeze temperature is preferably -80 DEG C, and freeze-drying time is preferably for 24 hours；
The single-layer tubular small intestine acellular matrix film that the sterilizing will preferably be sealed is through linear accelerating with 8000 ladd radiological doses
Radiation sterilization.
The present invention provides above-mentioned preparation method, the available good natural tubulose small intestine acellular matrix of this method
Film.
The following examples are intended to illustrate the invention, but is not used in and limits the scope of the invention, unless otherwise specified, this implementation
The conventional means that technological means used in example is well known to those skilled in the art, raw materials used is commercial goods.
The preparation of the natural tubulose ECM of single layer
The white pig of real estate 1 is taken, weight continues under ether inhalation anesthesia in 50-100Kg, chooses midline abdominal incision, stringent standby
Skin, disinfection spread aseptic towel list, and cutting closer intercepts chitterlings, and every segment length is 15 centimetres, immerse 4 DEG C of physiology after taking-up immediately
In salt water, and small intestine is cleaned with physiological saline.Small intestine after cleaning up is nested on annular tube shaped bracket, is destroyed with scissors
Placenta percreta, it is as much as possible uniformly to cut off tunica serosa intestini tenuis layer, then carried out with tunica serosa intestini tenuis layer of the poly (methyl methacrylate) plate to destruction
It strikes off.After the completion of striking off, small intestine is overturn, then strikes off the mucous layer, submucosa and muscle layer of small intestine with same method, to small intestine basal cell
It is removed.Above each layer is carefully struck off, to prevent cell residue, obtains single-layer tubular small intestine acellular matrix film.To the pipe
Shape small intestine acellular matrix film is dried in vacuo after carrying out disinfection.
Fig. 1 is the photo figure before the natural tubulose chitterlings acellular matrix film inflation of single layer of the present invention；Fig. 2 is that the present invention is single
Photo figure after the natural tubulose chitterlings acellular matrix film inflation of layer；Fig. 3 is that the natural tubulose chitterlings of single layer of the present invention are de- thin
Photo figure after the vacuum drying of cytoplasmic matrix film；Fig. 1 to Fig. 3 illustrates that after the natural tubulose ECM inflation, tube wall surrounding is uniformly complete,
Without breakage, it was demonstrated that natural tubulose small intestine acellular matrix film has successfully been prepared in this method.
The natural tubulose chitterlings acellular matrix film for the single layer that Fig. 4 is prepared for the present invention is put to be swept under 1000 times of Electronic Speculum
Retouch photo；The natural tubulose chitterlings acellular matrix film for the single layer that Fig. 5 is prepared for the present invention is put to be swept under 5000 times of Electronic Speculum
Retouch photo.Figure 4 and 5 explanation, the tubulose ECM inner cell produced of the present invention strike off it is very clean thoroughly, can be with by this method
Good natural tubulose chitterlings acellular matrix film is made.
Claims (6)
1. a kind of preparation method of natural tubulose small intestine acellular matrix film, which is characterized in that this method comprises:
Step 1: the small intestine intercepted in animals is immersed in physiological saline and is cleaned, the small intestine after cleaning up is set
Enter closed container, be rapidly frozen in -80 DEG C of liquid nitrogen, it is spare that defrosting is taken out after half an hour；
Step 2: the small intestine after defrosting is nested on annular tube shaped bracket, is destroyed placenta percreta with scissors, is cut off tunica serosa intestini tenuis
Layer, is then struck off with tunica serosa intestini tenuis layer of the poly (methyl methacrylate) plate to destruction, and mucous layer, the mucous membrane of small intestine are struck off after overturning small intestine
Lower layer and muscle layer, to small intestine, basal cell is removed, and obtains single-layer tubular small intestine acellular matrix film；
Step 3: the single-layer tubular small intestine acellular matrix film that step 2 obtains is subjected to freeze-drying sealing, sterilizing preservation, obtains list
The natural tubulose small intestine acellular matrix film of layer.
2. a kind of preparation method of natural tubulose small intestine acellular matrix film according to claim 1, which is characterized in that institute
The animal of the step of stating one is pig, ox, sheep, mouse, horse.
3. a kind of preparation method of natural tubulose small intestine acellular matrix film according to claim 1, which is characterized in that institute
The cleaning of the step of stating one is cleaned in 4 DEG C of physiological saline.
4. a kind of preparation method of natural tubulose small intestine acellular matrix film according to claim 1, which is characterized in that will
Small intestine after cleaning up is placed in closed container, is rapidly frozen in -80 DEG C of liquid nitrogen, takes out and thaws after half an hour.
5. a kind of preparation method of natural tubulose small intestine acellular matrix film according to claim 1, which is characterized in that institute
Four freeze temperature of the step of stating is -80 DEG C, and freeze-drying time is for 24 hours.
6. a kind of preparation method of natural tubulose small intestine acellular matrix film according to claim 1, which is characterized in that institute
The sterilizing of the step of stating four is that the multilayer tubular small intestine acellular matrix film that will be sealed is radiated through linear accelerating with 8000 ladds
Dosage radiation sterilization.
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| CN201810839013.6A CN108939157A (en) | 2018-07-27 | 2018-07-27 | The preparation method of natural tubulose small intestine acellular matrix film |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108939160A (en) * | 2018-07-27 | 2018-12-07 | 吉林大学 | A kind of artificial tubular biomaterial |
| CN113151177A (en) * | 2021-05-21 | 2021-07-23 | 四川大学华西医院 | Mammary gland or breast cancer tissue acellular matrix and preparation method and application thereof |
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|---|---|---|---|---|
| US20030219417A1 (en) * | 1999-11-22 | 2003-11-27 | Bioscience Consultants, Llc | Bioreactor mediated recellularization of natural and tissue engineered vascular grafts |
| CN101366975A (en) * | 2008-09-03 | 2009-02-18 | 陕西瑞盛生物科技有限公司 | Preparation method for cellfree intestinum tenue submucosa biological material |
| CN103272277A (en) * | 2013-05-28 | 2013-09-04 | 北京博辉瑞进生物科技有限公司 | Mold for preparing accellular small intestine submucosa matrix material |
| CN108310464A (en) * | 2018-03-21 | 2018-07-24 | 吉林大学 | Natural tubulose small intestine acellular matrix film and preparation method thereof |
-
2018
- 2018-07-27 CN CN201810839013.6A patent/CN108939157A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030219417A1 (en) * | 1999-11-22 | 2003-11-27 | Bioscience Consultants, Llc | Bioreactor mediated recellularization of natural and tissue engineered vascular grafts |
| CN101366975A (en) * | 2008-09-03 | 2009-02-18 | 陕西瑞盛生物科技有限公司 | Preparation method for cellfree intestinum tenue submucosa biological material |
| CN103272277A (en) * | 2013-05-28 | 2013-09-04 | 北京博辉瑞进生物科技有限公司 | Mold for preparing accellular small intestine submucosa matrix material |
| CN108310464A (en) * | 2018-03-21 | 2018-07-24 | 吉林大学 | Natural tubulose small intestine acellular matrix film and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108939160A (en) * | 2018-07-27 | 2018-12-07 | 吉林大学 | A kind of artificial tubular biomaterial |
| CN113151177A (en) * | 2021-05-21 | 2021-07-23 | 四川大学华西医院 | Mammary gland or breast cancer tissue acellular matrix and preparation method and application thereof |
| CN113151177B (en) * | 2021-05-21 | 2023-11-03 | 四川大学华西医院 | Breast or breast cancer tissue acellular matrix and preparation method and application thereof |
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