CN1297324C - Tissue engineering autologous complex skin and its preparation method - Google Patents
Tissue engineering autologous complex skin and its preparation method Download PDFInfo
- Publication number
- CN1297324C CN1297324C CNB03150373XA CN03150373A CN1297324C CN 1297324 C CN1297324 C CN 1297324C CN B03150373X A CNB03150373X A CN B03150373XA CN 03150373 A CN03150373 A CN 03150373A CN 1297324 C CN1297324 C CN 1297324C
- Authority
- CN
- China
- Prior art keywords
- skin
- cell
- patient
- autologous
- bluk recombination
- 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.)
- Expired - Lifetime
Links
Images
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The present invention discloses a tissue engineering autologous complex skin which is composed of an autologous dermal cell layer and an autologous hypodermal cell layer attached to a fibrin biologic bracket. The preparation method of the autologous complex skin comprises the following steps: an epidermal stem cell and a dermal fibroblast from a patient self are amplified and differentiated in vitro; then planting the epidermal cell on a fibrin biologic bracket with the dermal fibroblast of the patient self for culture; constructing the tissue engineering autologous complex skin in vitro. The tissue engineering autologous complex skin is applied to the preparation of biomaterials for skin transplantation. The present invention has the advantages: 1, the present invention is constructed by autologous skin cells and has no rejection reaction, and the tissue re-formed from the cells has long survival time; 2, the adopted biologic bracket has good adhesiveness, high survival rate after transplantation and quick wound surface restoration; 3, the present invention has the epithelial cell and the fibroblast forming dermal tissue, the healed skin is hard, has little possibility of break and has the advantages of good healing effect and normal appearance.
Description
Technical field
The present invention relates to a kind of organizational project from bluk recombination skin and preparation method thereof, belong to field of tissue engineering technology.
Background technology
Tissue engineering technique is a kind of new and high technology that is used to prepare transplantable, as to possess morphological characteristic and function tissue and organ of setting up behind one nine eight zero years in the world, the product of researching and developing in the world at present has: skin, corium, mucosa, tendon, bone, cartilage, blood vessel, nerve, cornea, cardiac valve, kidney, body of gland and digestive appartus official rank, and achieving success at first is skin histology, dermal tissue and soft tissue.Skin and dermal tissue are to obtain the product that U.S. FDA authenticates prior to 1999, comprising Apligraf, Integraf and three kinds of products of Dermagraf.Because organizing of tissue engineering technique preparation includes cell alive and possible Biodegradable material in the organ pipe, product can be used for body surface or transplants in body, Given this series products biological product different from the past, chemical drugs, apparatus etc., U.S. FDA has been set up the new group that examines specially in verification process, the place of examining takes the lead by apparatus, examines with weapon exercises.The organization engineering skin that the homogeneous variant cell of drugs approved by FDA is produced has clinical effectiveness preferably in a short time, but still has the problem of three aspects in clinical use: the first, because the rejection of allosome makes its time to live short; The second, its support absorbs and is unfavorable for wound healing slowly; The 3rd, owing to be independent epithelial cell, the skin after the healing is very thin, and is easily broken, causes new wound.
Summary of the invention
The purpose of this invention is to provide a kind of organizational project from bluk recombination skin.
Another object of the present invention provides the preparation method of organizational project from bluk recombination skin.
The 3rd purpose of the present invention provides the application of this organizational project from bluk recombination skin.
For achieving the above object, the present invention is by the following technical solutions:
A kind of organizational project is from bluk recombination skin, and this composite skin is by being attached to the auto derma cellular layer on the fibrin biological support and constituting from the body surface skin cell layer.
Described auto derma cellular layer is to obtain through amplification in vitro, cultivation deriving from the dermal fibroblast of patient from body.
Described is to obtain through amplification in vitro, cultivation deriving from the epidermal stem cells of patient from body from the body surface skin cell layer.Epidermal stem cells divides formation epidermal stem cells and TA cell (transientamplifying cell) in incubation, wherein epidermal stem cells has multiplication capacity, and the TA cell has differentiation capability, and differentiation becomes epithelial cell, constitutes epidermal area.
The fibrin that described fibrin is behaved.The advantage of this support is: the first, the amplification of energy support matrix chrotoplast and hypodermal cell; The second, this support can be absorbed faster; The 3rd, its support softness, but wrinkle, operation easily, the 4th, stronger adhesiveness is arranged, be easy to be attached to wound face.
A kind of organizational project is from the preparation method of bluk recombination skin, be will derive from the patient from the epidermal stem cells of body and dermal fibroblast in amplification in vitro and differentiation, to plant behind the epidermis cell again in containing on the fibroblastic fibrin biological support of patient's auto derma, cultivate, in the external structure organizational project from bluk recombination skin.
The construction method of the fibroblastic fibrin biological support of the described patient's of containing auto derma is: adopting people's Fibrinogen is material, is 1-8mg/ml according to concentration, adds people's thrombin 1-5unit/ml and CaCl
21-5 μ mol/ml adds dermal fibroblast simultaneously, mixes under the room temperature, is implemented at once on the culture dish; Add DMEM and 10%FBS culture medium culturing.
Described plantation is meant and is built with in the culture dish of biological support to fibroblastic amount adding of having used with 10 times deriving from the epidermis cell of patient from body.
The quantity of described adding dermal fibroblast is 40000-60000 cell/ml.
Described organizational project is used to prepare the biomaterial that skin transplantation is used from bluk recombination skin.
Organizational project adopts from body epithelial cell and hypodermal cell behind amplification in vitro from bluk recombination skin, is inoculated on the degradable dermal scaffold and cultivates, and at the external structure organization engineering skin, itself and natural skin have similar 26S Proteasome Structure and Function.Be transplanted to patient from body then, be used for large-area burns patient and skin ulcer patient's skin transplantation and reparation.This goods organizational structure is similar to or is equal to people's skin histology, adopts patient's autogenous cell, does not have the cross infection problem of immunological rejection and infectious disease, is to treat the very effective method of skin burn patient at present.Can be widely used in treating the skin injury that multiple reasons such as skin burn, ulcer, wound cause, especially can be used for the first aid of large area skin burn.
The present invention can use for transplanting at short notice for the patient provides large-area epidermis and hypodermal cell, and its advantage is:
1, is the autologous skin cell construction, thereby do not have rejection that cell forms again, and to be organized into live time long.
2, the biological support adhesiveness of Cai Yonging is good, transplants back survival rate height at epithelial cell and hypodermal cell, and the wound face reparation is fast.
3, the existing epithelial cell of this biological engineering composite skin the fibroblast that constitutes its dermal tissue is arranged again, thereby its skin wound healing skin is solid, is difficult for fragmentation, and healing effect is good, and outward appearance is more normal.
The invention will be further described below in conjunction with the drawings and specific embodiments.
Description of drawings
Fig. 1 is a finished product appearing diagram of the present invention
Fig. 2 carries out local skin condition diagram before the skin transplantation for the patient
Fig. 3 is the local skin condition diagram after the skin transplantation
The specific embodiment
Before definite bark fetching skin tissue, by medical worker with qualifications of a licensed doctor to patient or family numbers of patients explanation reasons, strive to such an extent that patient or family members agree, the position of drawing materials of skin is determined by the medical worker with qualifications of a licensed doctor, and is drawn materials according to the operation principle of hospital's regulation.The patient checks UP and chemically examines, and comprises the inspection of routine inspection, infectious disease cause of disease (as HIV, hepatitis virus etc.).The skin of being got is put into sterile chamber by the professional immediately, and cold preservation immediately (below 10 ℃), and S.O.P. is in accordance with regulations handled then.
The preparation of embodiment 1, tissue engineering composite skin
One. the patient is gone down to posterity from the extraction and the cell culture of body healthy cell
(1) patient is from the extraction of body healthy cell
1. operating room is drawn materials
1) with medical iodine tincture cleaning patient bark fetching face;
2) with normal saline flushing bark fetching face, remove residual iodine tincture;
3) get the long narrow skin of 1cm * 2.5cm;
4) skin that takes off is put and is sent cell culture chamber in the stock solution.
2. the preservation of skin histology after drawing materials: DMEM (purchasing the Hyclone company in the U.S.) culture fluid that contains the streptomycin of 100 units/ml penicillin and 100mg/ml with 5ml is preserved tissue (4 ℃) in preserving pipe.
3. sterilization in superclean bench, cleaning organizes bulk to separate: with the 0.1M PBS flushing tissue of the streptomycin that contains 100 units/ml penicillin and 100mg/ml, to prune unnecessary fat and subcutaneous knot a kind of thick silk tissue.Tissue is cut into the piece of tissue of 1cm * 1cm.
4. tissue digestion (Sony CO
2Incubator): 3～4 1cm * 1cm epidermal tissue piece and dermal tissue piece be positioned over respectively to add 10ml concentration in the 60mm culture dish be pancreatin/EDTA liquid of 0.25%, 37 ℃, 120 minutes; The DMEM+10%FBS culture fluid stops.
5. cell separation: postdigestive piece of tissue is shredded and breaks up with suction pipe.
6. centrifugal: the cell harvesting after dispelling to centrifuge tube, Backman company temperature control centrifuge, 25 ℃, 1000 commentaries on classics/min, centrifugal 5 minutes, abandoning supernatant.
7. cell is identified
1) morphology: adopt and be inverted observation by light microscope.Epithelial cell is multiangular more, and the angle is the obtuse angle, and cell is round property; The cell appearance transparent, cell is tightly linked into monolayer, also can grow in a tubular form.The different projection of the protruding 2-3 of a dermal fibroblast matter length is polygon or long prismatic.
2) cell phenotype: adopt keratoprotein antibody and collagen antibody (American I CN company) to carry out immunizing antigen and identify, the cell that obtains of display separation is normal epidermis cell and normal dermal fibroblast as a result.
3) carcinogenecity: adopt the nude mice test, see " the consideration main points that U.S. FDA is identified and accused with cell strain about biological product production ", the result shows non-carcinogenesis.
4) aseptic detection: undertaken by " Chinese biological goods rules " (2000 editions) general rule " biological product sterility test rules " A/B item, the result meets aseptic requirement.
5) cell survival rate: adopt trypan blue (Trypan blue) staining inspection, the cell of nuclear staining is a dead cell, and non-staining cell is a living cells, counts total cell number and dead cell number, calculates cell survival rate.
Cell survival rate=[(total cell number-dead cell number)/total cell number] * 100%
After measured, survival rate surpasses 80%.
(2) epidermis cell former is commissioned to train and supports and go down to posterity
1. primitive cell culture: add 10ml serum-free epithelial cell culture fluid (Sigma company, the U.S.) and blow outstandingly in centrifuge tube, 4 * 60mm culture dish is gone in inoculation; Sony CO
2Incubator, 37 ℃, 5%CO
2, cultivate.
2. passage cell is cultivated: changed liquid 1 time every 2～3 days, epithelial cell reaches and goes down to posterity after 50-70% compiles cell.
(3) hypodermal cell former is commissioned to train and supports and go down to posterity
1. primitive cell culture: add 10ml serum-free epithelial cell culture fluid (Sigma company, the U.S.) and blow outstandingly in centrifuge tube, 4 * 60mm culture dish is gone in inoculation; Sony CO
2Incubator, 37 ℃, 5%CO
2, cultivate.
2. passage cell is cultivated: changed liquid 1 time every 2～3 days, epithelial cell reaches and goes down to posterity after 50-70% compiles cell.
Two, make up the biological engineering composite skin:
(1) structure of biological support
1. prepare the human fibrinogen
With 200ml blood plasma 4 ℃ centrifugal, 3500 commentaries on classics/min, 30 minutes, abandon supernatant, add 10ml water for injection, make suspension, preserve standby.
2. make up and be attached with the fibroblastic biological support of auto derma
Adopting people's Fibrinogen is material, according to concentration is 1-8mg/ml, thrombin (the purchasing company) 1-5unit/ml and the calcium ion 1-5 μ mol/ml that add the people in Sigma, the fibroblast that adds 50000 cells/ml simultaneously, indoor relaxing the bowels with purgatives of warm nature mixes, be implemented at once on the culture dish of 60mm or 100mm, form and be attached with the fibroblastic biological support of auto derma.Add DMEM and 10%FBS culture medium culturing, fibroblast proliferation forms the hypodermal cell layer.Because timbering material has adhesiveness, can help being adsorbed onto wound surface.
3. structure epithelium layer
1) plantation: the epithelial cell of amplification cultivation is built with in the culture dish of biological support to fibroblastic amount adding of having used with 10 times, and epithelial cell will be planted in the rack surface that contains the hypodermal cell layer.
2) continue to cultivate: continue to cultivate with DMEM and epithelial cell culture fluid (Sigma company), form epidermal area, 37 ℃, 5%CO
2, get final product clinical practice after 24 hours.
(2) biological support and finished product detection
1. the detection of biological support
1) pH value is measured: the support 5g that draws materials, and the distilled water 15ml of adding pH7.0 soaked 24 hours down in 37 ℃, surveyed pH, and pH value is between 7.0-7.5.
2) external degradation test: the support of drawing materials, immerse in 37 ℃ of phosphate buffers that contain protease (2mg/ml), soak after 24 hours, support all disappears.
3) skin irritation and sensitization test (STT): by GB/T16886.10-2000 medical apparatus and instruments biological assessment the 10th part: stimulate with the sensitization test (STT) regulation and test, the result does not have sensitization.
4) cytotoxicity is according to GB/T16886.5-1997 medical apparatus and instruments biological assessment the 5th part: the regulation of cell toxicity test is tested, as a result no cytotoxicity.
5) muscle implantation test is by GB/T16886.6-1997 medical apparatus and instruments biological assessment the 6th part: the method for implanting regulation in the local response test of back is carried out, and the result does not have local response.
6) genetic toxicity test is tested according to the method for stipulating in GB/T16886.3-1997 medical apparatus and instruments biological assessment the 3rd part, as a result avirulence.
2. finished product detection
1) outward appearance: it is membranaceous to be translucent, and thickness is even, glossy, sees Fig. 1.
2) specification: the size dimension that is not less than sign.
3) elasticity and requirement of strength: artificial skin should be able to recover its initial and design shape during folding and distortion, is kept perfectly.
4) histological observation: the organizational project holostrome skin of cultivating is carried out conventional organization learn embedding treatment, 10% formalin fixed, organized processing, paraffin embedding are carried out H-E dyeing, observation by light microscope after the mounting after the section.Result: under the light microscopic, do not see tangible bacterial growth, do not have tangible epithelium or fibroblast regression or necrosis, the random growth of no locality cell; As seen the organization engineering skin of In vitro culture has epithelial layer and skin corium, and wherein epithelial layer has a large amount of epithelial cells, and skin corium has a large amount of hypodermal cell.
5) sterility test: undertaken by " Chinese biological goods rules " (2000 editions) general rule " biological product sterility test rules " A/B item, the result meets aseptic requirement.
The using method of product: before the use, with the culture fluid flushing that does not contain serum three times.When transplanting, with aseptic scalpel or surgery tweezers necrotic tissue cleaning with wound surface, hemostasis, then with normal saline towards Xian Sanci, organizational project is spread on wound surface from bluk recombination skin, and sutured around making is covered in organizational project on bluk recombination skin with oily sand, use certain pressure to make its new subsides, fixing with the cotton yarn packing then.Open parcel after 5-7 days, visible newborn skin (red, pink) growth, wound heals fully after one month.
The storage of product and transportation should be carried out in 2 ℃～10 ℃, shady and cool, dry, the environment that cleans, avoid sunlight direct projection, non-corrosiveness gas, no weight.
Embodiment 2, clinical report
One. include the case standard in
1. the deep burn patient cuts, cuts wound surface after the crust.
2. the ulcer wound surface of the long-term disunion that causes of a variety of causes and granulation wound (intractable ulcer wound surface such as III degree burn back granulation wound in late period, diabetic foot etc.).
3. the extensive scar hyperplasia of whole body, contracture deformity, or keloid, hypertrophic cicatrix, cicatrix is hard, itch, bitterly, needs excision cicatrix skin-grafting person.
Belong to above-mentioned situation patient, and the age is no more than 60 years old for including object in.
Two. get rid of the case standard
1. complex injury such as shock stage, chemical poisoning.
2. merge serious primary disease such as cardiovascular, cerebrovascular, liver, kidney, hemopoietic system.
3. allergic constitution person or autoimmune disease person.
Three. draw materials
1. check blood, routine urinalysis, liver, renal function before the art.
2. the art of drawing materials: look the wound surface size, get a place or several places holostrome skin of normal skin 1.5cm * 0.5cm size under local anaesthesia, skin donor site is directly sewed up with the 3-0 silk thread.Skin histology places preserves liquid.
Four. the laboratory In vitro culture
Place immediately after skin histology is drawn materials and preserve liquid, be placed in 2 ℃-10 ℃ the couveuse transportation and preserve, in 6 hours, cultivate, be that carrier is cultivated the autologous skin stem cell with the bioengineered tissue, and meet its organizational project from bluk recombination skin Registering product standard.
Five. reject the case standard
Skin progenitor cell is cultivated failure, and the organizational project of cultivating gained does not meet product standard person from bluk recombination skin.
Six. transplant operation
Composite skin art: after 2～3 weeks, the composite skin of cultured autologous skin stem cell, hypodermal cell and biological support is transplanted to wound surface, granulation wound, ulcer wound surface and scar excision after the crust cut, cut to deep burn, wound surface after loosening.
1. wound surface should be fresh, and blood fortune is arranged, and no slough does not have and infects, and wound surface is removed thoroughly, a large amount of antibiotics normal saline flushings, and hemostasis is thoroughly.
2. the composite skin that will be attached on the kpetrolatum gauze is transplanted on wound surface, and corium faces down, and epidermis side upwards, with the 3-0 silk thread composite skin is sewed up together with the kpetrolatum gauze edge, be fixed in wound surface, stay long line, place the saline gauze with big mesh on kpetrolatum gauze, wrapping is pressed in packing a little.
3. be positioned at joint part as skin-grafting, art finishes fixed for two weeks with plaster slab.
Seven. post surgery treatment
Postoperative was with responsive antibiotics 5-7 days.
Granulation wound Composite skin postoperative 4-5 days, cut, cut after the crust behind wound surface or the scar excision wound surface 5-7 days, open outer dressing and cut off the packing suture, soak internal layer dressing with normal saline, after removing gently, inspect and transplant Composite Skin and be subjected to distinguish the situation that survives, color, secretions, the edge of wound situation of observing skin.Continue to press a little to pack with oil gauze, saline gauze and gauze, postoperative was changed dressings for 2 times weekly in one month, took out stitches in postoperative 7-10 days.
Eight. efficacy assessment standard
Produce effects: transplant month wound healing of composite skin more than 75%.
Effectively: transplant month wound healing of composite skin at 30%-75%.
Invalid: as to transplant month wound healing of composite skin and 30%.
Nine. data management and statistics
1. data collection:
All MethodsThe cases enrolled all must be finished the case observation table, and researcher will be observed, check result is timely, accurate, complete, standard, be recorded in case history and the case observation table truly.
After total data input database and the check and correction, hand over the statistical analysis personnel to carry out statistical analysis the data base, and statistical report is provided, hand over the main researcher of clinical trial to write out the clinical summary report.
2. statistical analysis
According to data character, to basic data data, efficacy analysis (each index analysis) and safety analysis, carry out descriptive statistics respectively.
The continuous data statistical method: situation of change adopts paired sample t check before and after in the observation group.
Enumeration data statistical method: adopt definite probabilistic method.
Grade type data statistical approach: situation of change symbolization rank test before and after in the observation group.
3, statistical software: all statistics all utilize SAS JMP5.0 software to carry out statistical analysis, adopt two-sided test, with P≤0.05 as statistical significance is arranged.
Ten. conclusion
This product has carried out clinical research in Beijing liang man front three hospital, and selected altogether 20 routine patients observed through postoperative in one month, estimated it and transplanted the improvement situation of repairing deep burn and cicatrix for tissue engineering composite skin, and the curative effect of each MAIN OUTCOME MEASURES is as follows:
(1) wound secretion: all do not have wound surface secretions and ooze out.
(2) wound surface swelling: all the noinvasive swelling of the face expands.
(3) edge of wound reaction: be negative reaction (promptly not having the edge of wound reaction)
(4) wound healing percentage rate: (being produce effects) 15 examples: 30%-75% (effectively) 1 example more than 75%, (invalid) below 30% 4 examples.
Postoperative one month: total effectively case 16 examples, total effective rate 80%.
This Figure of description exemplify before patient's art and the situation map of postoperative to explain, see Fig. 2 and Fig. 3 respectively, as can be seen at the autodermic position of operation transplantation organizational project of the present invention, newborn skin easily healing is effective, outward appearance is normal, and patient's symptom is improved.
Claims (8)
1. an organizational project is characterized in that from bluk recombination skin: this composite skin constitutes by the auto derma cellular layer on the fibrin biological support that is attached to the people with from the body surface skin cell layer.
2. organizational project according to claim 1 is characterized in that from bluk recombination skin: described auto derma cellular layer is to obtain through amplification in vitro, cultivation deriving from the dermal fibroblast of patient from body.
3. organizational project according to claim 1 is characterized in that from bluk recombination skin: described is to obtain through amplification in vitro, cultivation deriving from the epidermal stem cells of patient from body from the body surface skin cell layer.
4. an organizational project is from the preparation method of bluk recombination skin, be will derive from the patient from the epidermal stem cells of body and dermal fibroblast in amplification in vitro and differentiation, to plant behind the epidermis cell on the fibrin biological support that contains the fibroblastic people of patient's auto derma again, cultivate, in the external structure organizational project from bluk recombination skin.
5. organizational project according to claim 4 is from the preparation method of bluk recombination skin, it is characterized in that: the construction method of the fibroblastic fibrin biological support of the described patient's of containing auto derma is: adopting people's Fibrinogen is material, according to concentration is 1-8mg/ml, adds people's thrombin 1-5unit/ml and CaCl
21-5 μ mol/ml adds dermal fibroblast simultaneously, mixes under the room temperature, is implemented at once on the culture dish; Add DMEM and 10%FBS culture medium culturing.
6. organizational project according to claim 4 is characterized in that from the preparation method of bluk recombination skin: described plantation is meant and is built with in the culture dish of biological support to fibroblastic amount adding of having used with 10 times deriving from the epidermis cell of patient from body.
7. organizational project according to claim 5 is characterized in that from the preparation method of bluk recombination skin: the quantity of described adding dermal fibroblast is 40000-60000 cell/ml.
8. claim 1 or 2 or 3 described organizational projects are used to prepare the biomaterial that skin transplantation is used from bluk recombination skin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB03150373XA CN1297324C (en) | 2003-07-25 | 2003-07-25 | Tissue engineering autologous complex skin and its preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB03150373XA CN1297324C (en) | 2003-07-25 | 2003-07-25 | Tissue engineering autologous complex skin and its preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1569258A CN1569258A (en) | 2005-01-26 |
| CN1297324C true CN1297324C (en) | 2007-01-31 |
Family
ID=34472711
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB03150373XA Expired - Lifetime CN1297324C (en) | 2003-07-25 | 2003-07-25 | Tissue engineering autologous complex skin and its preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1297324C (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1974015A4 (en) * | 2006-01-27 | 2012-07-04 | Univ California | Biomimetic scaffolds |
| CN101143231B (en) * | 2007-10-19 | 2011-05-18 | 中国人民解放军第四军医大学 | Tissue engineering skin containing muscle cell and preparation method thereof |
| CN101297981B (en) * | 2008-07-03 | 2011-11-02 | 中国检验检疫科学研究院 | Tissue engineering skin construction method and tissue engineering skin produced by the same |
| CN102091352A (en) * | 2009-12-09 | 2011-06-15 | 中国人民解放军总医院第一附属医院 | Method for constructing tissue engineering skin model containing sweat gland |
| CN102462864B (en) * | 2010-11-12 | 2014-06-18 | 中国辐射防护研究院 | Novel method for constructing tissue engineering skin |
| CN102488930A (en) * | 2011-12-15 | 2012-06-13 | 无锡市第三人民医院 | Method for preparing auto and allo-epidermal cell mixed suspension |
| CN104399125B (en) * | 2014-12-01 | 2016-03-16 | 中国人民解放军第三军医大学第三附属医院 | The method that epidermal stem cells breaks up to sweat gland sample epithelial cell |
| CN105176914A (en) * | 2015-10-13 | 2015-12-23 | 怡诺博(北京)生物医学技术有限公司 | Simultaneous autologous epidermic cell and fibrocyte preparation method and biological cosmetic product thereof |
| CN110075126A (en) * | 2019-05-08 | 2019-08-02 | 张永国 | A kind of gingival cell hydrogel filler and preparation method thereof, application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1198090A (en) * | 1995-07-28 | 1998-11-04 | 埃索拉根技术有限公司 | Use of autologous dermal fibroblasts for repair of skin and soft tissue defects |
| CN1310027A (en) * | 2001-02-06 | 2001-08-29 | 中国人民解放军第三军医大学第三附属医院 | Preparation of compound artificial skin |
| CN1363398A (en) * | 2002-01-16 | 2002-08-14 | 吉林大学 | Stratified artificial skin using chitosan or its derivative as matrix clathrum |
-
2003
- 2003-07-25 CN CNB03150373XA patent/CN1297324C/en not_active Expired - Lifetime
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1198090A (en) * | 1995-07-28 | 1998-11-04 | 埃索拉根技术有限公司 | Use of autologous dermal fibroblasts for repair of skin and soft tissue defects |
| CN1310027A (en) * | 2001-02-06 | 2001-08-29 | 中国人民解放军第三军医大学第三附属医院 | Preparation of compound artificial skin |
| CN1363398A (en) * | 2002-01-16 | 2002-08-14 | 吉林大学 | Stratified artificial skin using chitosan or its derivative as matrix clathrum |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1569258A (en) | 2005-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107281550B (en) | Preparation method of co-crosslinked double-network hydrogel scaffold for promoting cartilage injury repair | |
| JP4615223B2 (en) | Collagen biofiber and its preparation method and use | |
| CN1291761C (en) | Closed cell culture system | |
| CN104055795B (en) | A kind of injectable implant and preparation method thereof | |
| CN1775189A (en) | Acellular dermal matrix and preparation method thereof | |
| CN107029296A (en) | Periosteum patch, the preparation method and application of a kind of Guided Bone Regeneration | |
| CN101002964A (en) | Tissue engineering composite skin material, and its preparing method | |
| CN1668732A (en) | Treatments with autologous fibroblast | |
| CN107073168A (en) | Tissue grafts | |
| CN1297324C (en) | Tissue engineering autologous complex skin and its preparation method | |
| CN1927413A (en) | Amnion stroma tectorial blood vessel internal branch and preparing method thereof | |
| CN108057116A (en) | Application of the stem cell composition in skin injury medicine | |
| CN107890586B (en) | Preparation method of allogeneic biological breast patch | |
| JP2019529015A (en) | Dermal layer for transplantation with increased survival rate and method for producing the same | |
| CN1618954A (en) | Bioderived amniotic membrane, composite bioderived amniotic membrane and preparation method thereof | |
| CN1799558A (en) | Glue adhesion amnion and preparation method thereof | |
| CN108066750A (en) | Stem cell and its secretion are used to treat the new application of skin burn | |
| CN101306208A (en) | Human liquid state derma preparation for injection and its preparation method | |
| CN1289156C (en) | Tissue engineering autologous cornea epithelium and its preparation method | |
| CN1916166B (en) | Method for preparing epithelium of autologous cornea | |
| CN1234429C (en) | Artificial skin, preparing method and application thereof | |
| CN110433340B (en) | Acellular matrix urethral suspension repair material and preparation method and application thereof | |
| CN107224618A (en) | Promote active scaffold material, its preparation method and the application of osteanagenesis | |
| CN109072185A (en) | Enhanced pluripotent cell and microvascular tissue and its application method | |
| CN1062441C (en) | composite living skin equivalents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: LV WEIGUANG Free format text: FORMER OWNER: BEIJING YILING BIOENGINEERING CO., LTD. Effective date: 20060602 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20060602 Address after: 100022, No. 1707, Jianguo Road, No. 98, Jianguo Road, Beijing, Chaoyang District, A Applicant after: Lv Weiguang Address before: 101500 Beijing City Industrial Development Zone in Miyun Dashenglu No. 60 Applicant before: Beijing Yiling Bioengineering Co.,Ltd. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term |
Granted publication date: 20070131 |
|
| CX01 | Expiry of patent term |