CN101361990B - Double layer artificial skin and preparation method thereof - Google Patents
Double layer artificial skin and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a double-layer artificial skin and a preparation method thereof, wherein, cell-free membranous biological derivative material is used as a surface layer, and a fibroblast, extracellular matrix synthesized and secreted by the fibroblast and a cell growth factor are compounded in the interior of biological support material to form a dermis, and then the surface layer and the dermis are combined to form the double-layer artificial skin; a compact surface layer structure can effectively reduce the loss of water, electrolytes and protein from surface of wound, avoid the invading and the reproduction of bacteria to the impaired surface of wound as well as the infection of the surface of wound, and be beneficial to epitheliosis and epithelial growth; the dermis can directly repair the surface of wound, promote the ingrowth of cells around the surface of wound and the angiogenesis, induce the differentiation from stem cells to skin cells and quicken wound healing; the artificial skin has the advantages of being capable of promoting the regeneration of skin, improving the elasticity, the flexibility and the mechanical abrasion resistance of skin after the surface of wound is healed, reducing excess scar tissues, controlling the contracture, having excellent biocompatibility, increasing the success rate of transplant and improving the quality of healing; the double-layer artificial skin has wide material resources and simple production method, and is applicable to the clinical treatment of skin defect caused by inflammation, ulcer, thermal burns, iatrogenicity and the like.
Description
Technical field
The invention belongs to the technical field of biological materials of organizational project, be specifically related to a kind of double-layered artificial skin and preparation method thereof.
Background technology
Skin is the vital tissue organ that covers and protect body surface.Owing to reasons such as inflammation, ulcer, wound, burn, tumor post-operation and congenital abnormality often cause the damaged and unusual of skin.For the at present methods that adopt auto-skin grafting of this tissue defect, this not only causes the new wound defective of skin donor site, and often receives the restriction that supplies the skin source more.
Along with the development of tissue engineering technique, the preparation artificial skin has become the focus of skin injury treatment.The ideal tissue engineering skin can imitate the structure of natural skin, has the compound skin of epidermal area and skin corium.The skin corium composition wherein and the composition of dermis of skin are approaching, are beneficial to growing into of dermal cell; The epidermal area coverture plays the isolation external environment, prevents moisture evaporation, avoids bacterial invasion, and more compound growth factors or medicine can play the purpose that promotes wound healing, treatment dermatosis simultaneously.
Commercial at present double-layered artificial skin has Integra (production of Integra LifeSciences company); Be the artificial skin of double-deck matrix, the corium part is processed the support with certain hole size by the collagen protein and the CHS of ox tendon, and epidermal area then is made up of pellosil; Can control evapotranspiring of wound aqueous vapor; At wound healing process, skin corium can combine with wound, and the pellosil of epidermal area can be removed in skin corium healing back; Further do auto-skin grafting again, U.S. FDA was checked and approved Integra and has been used in the burn and scald treatment in 1996; Its deficiency mainly is, because there is nondegradable glued membrane on its surface, is difficult for observing the early stage infection conditions of the surface of a wound, is unfavorable for the sealing with the surface of a wound of creeping of edge of wound epidermic cell, and the later stage is taken pellosil off also needs second operation.Apligraf (production of Organogenesis company) also is double-deck artificial skin, and skin corium is to be planted in the bovine collagen albumen by human dermal fibroblast, and epidermal area is to be made up of human epidermal cell, attached to forming epidermal area on the corium; Its deficiency is that mainly epidermal area is to be made up of the higher allosome epidermic cell of immunogenicity, has increased the immunological rejection of product, makes complex manufacturing, PT length, cost increase simultaneously.
Summary of the invention
The object of the invention just provides a kind of double-layered artificial skin and preparation method thereof, has that preparation cycle is short, cost is low, method is easy, the advantage of easy handling; Prepared artificial skin has hemostasis, prevent that body fluid runs off, protects the surface of a wound and promotes the function of wound healing; Excellent biological compatibility is arranged; Can be used for used for reparing skin defect, control traumatic infection, the difficult more surface of a wound of property of treatment, also having shape, size and thickness simultaneously can be according to the advantage of demand preparation.
The double-layered artificial skin that the present invention proposes; Include inoblast; It is characterized in that: be by taking off the membranaceous bio-derived material of cell as the top layer; Extracellular matrix and ESC by inoblast and synthesis secretion thereof are compound in the inner skin corium, both chimeric formations of forming of biologic bracket material; Described to take off the membranaceous bio-derived material of cell be natural biological tissue after taking off cell and handling formed membranaceous bio-medical material, comprise take off cell submucous layer of small intestine, acellular dermal matrix, take off the cell manadesma, take off the cell submucous layer of bladder, take off the cell amnion, take off cell dural any; Described inoblast is the inoblast from body or allosome source; Maybe can be to the stem cell of inoblast differentiation; Comprise mesenchymal stem cells MSCs, fat mesenchymal stem cell, hemopoietic stem cell, skin progenitor cell, mescenchymal stem cell, muscle stem cell, any of liver stem cells, NSC; Described biologic bracket material is three-dimensional propping material to be provided for cell growth, comprises any or several kinds mixing of Fibrinogen, scleroproein, alginates, collagen protein, mucinase, CHS, chitosan, hydrogel, gelatin, POLYACTIC ACID, PGTA.
The preparation method of double-layered artificial skin of the present invention includes fibroblastic obtain, the take off preparation of cell biological derived material and the structure of double-layered artificial skin, and concrete steps are following:
Step 1). fibroblastic obtaining: the human fibroblasts carries out vitro culture by the cell culture processes of routine, and the extensive amplification of cell can adopt existent method to carry out;
Step 2). take off the preparation of cell biological derived material: will take from of the same race or xenogeneic dermal layer of the skin or manadesma or submucous layer of small intestine or amnion or dural any membranaceous biomaterial; Place and contain Peracetic Acid and alcoholic acid aqueous solution soaking disinfection was not less than 1 hour, clean with phosphate buffered saline buffer (PBS solution); If (adopt thicker biomaterial, like dermal layer of the skin, need place-80 ℃ more than freezing half a hour this moment; Guarantee that the material internal and external temperature reaches consistent, take out the back and thaw naturally that so multigelation is 2～5 times; Make the cell disintegration of breaking fully, clean) with PBS solution; The NaOH solution soaking that is placed on 0.1～1M is carried out degreasing, is taken off the cell processing; Embathe the neutrality to pH with PBS solution, it is inserted in the mixing solutions that contains DNA enzyme and alpha-galactosidase again, 37 ℃ of environment are handled more than 25 minutes down, remove residual DNA and α-gal natural antigen composition, reduce immunogenicity, clean with PBS solution; For it is combined with timbering material firmly; Adopt mechanical means evenly drilling on its material; Aperture 0.1～1mm, spacing 0.5～5mm takes off cell biological derived material and the foraminous of atresia the stack of cell biological derived material and compresses it is combined closely; Become bilayer behind the dry sterilization and take off the cell biological derived material, subsequent use;
Wherein, described Peracetic Acid and alcoholic acid volume final concentration are respectively 0.1～0.5% and 2～10%; The final concentration of said DNA enzyme and alpha-galactosidase is respectively 30～50U/ml and 10～25U/ml;
Step 3). the preparation of double-layered artificial skin nutrient solution; Its component by volume per-cent includes: commercial minimum must nutrient solution DMEM 67.5%, commercial nutrient solution F12 is 22.5%, foetal calf serum 5～15%, Prostatropin 2～100ng/ml, epithelical cell growth factor 2～100ng/ml, Regular Insulin 1～50ng/ml, HYDROCORTISONE INJECTIONS 10～500ng/ml, VITAMIN B4 0.2～0.25mM and Transferrins,iron complexes 1～10 μ g/ml;
Step 4). the structure of double-layered artificial skin: the gelating soln of preparation biologic bracket material; After the bilayer of preparation taken off the cell biological derived material and soak into gelating soln, place Procuring under 37 ℃ of environment; The inoblast of vitro culture is pressed 10
4～10
6The concentration of individual/ml is mixed in the gelating soln, again by 0.1～1ml/Cm
2The bilayer that is added drop-wise to Procuring takes off on porose of cell biological derived material, places 5%CO
2Solidify under 37 ℃ of conditions in the environment; Insert again cultivate 2～3 days in the bilayer skin nutrient solution after, be placed on and continue on the cultivation support of cultivating in the vessel to cultivate 5～8 days, change liquid every day between during cultivation, double-layered artificial skin is cultivated and is accomplished; Above-mentioned culture condition is 37 ℃, 5%CO
2Environment.
Wherein, the gelating soln of described biologic bracket material can be by following preparation: under 4 ℃ of conditions, press mass ratio with collagen 7～10: chitosan 0.5～1: mucinase 2～3: CHS 0.5～1 mixes; It is 6～20mg/ml solution that the acetic acid of use 0.1～0.5M is mixed with concentration with it; After the uviolizing, the foetal calf serum by its volume adding 10% adds the DMEM substratum again and makes its final concentration reach 10mg/ml under the ice bath; Regulate pH value to 7.2～7.4, process gelating soln.Also can adopt the formulated of other gelating solns.
The double-layered artificial skin that the present invention is prepared has top layer and skin corium bilayer structure, includes the viable cell from human body, can survive at the surface of a wound after being applied to the surface of a wound, can participate in the reparation of the surface of a wound directly; Taking off the cell biological derived material forms fine and close surface structure and can reduce moisture, ionogen and protein losing by the surface of a wound effectively; Stop invasion, the breeding of bacterium to the impaired surface of a wound; Prevent traumatic infection; Induce of the migration of the epidermic cell in wound week, help the propagation and the growth of epithelium to the surface of a wound; Skin corium and dermis of skin composition are approaching; The extracellular matrix and the various kinds of cell growth factor that include human body cell and synthesis secretion thereof; Direct wound repairing; Excellent biological compatibility can promote to create the generation, induced dry-cell of the growing into of pericyte, the blood vessel differentiation to skin cells, can obviously promote the healing of the surface of a wound.
Double-layered artificial skin of the present invention has certain elasticity and toughness, and big or small thickness can be controlled, and is with short production cycle, does not have obvious immunological rejection, has excellent biological compatibility.Its effect in clinical application has: as skin large defect patient's the surface of a wound covering and the covering of skin donor site; Repair malnutrition and infective wound surface; Treat the chronic difficulty ulcer surface of a wound of healing; The covering of postoperative wound surface and preventing infection; Promote growing into of surface of a wound granulation tissue, cell, promote the regeneration of damaged skin.
The double-layered artificial skin of the present invention's preparation is compared with present existing skin substitutes product, has the following advantages: can promote the regeneration of surface of a wound skin; Skin elasticity, snappiness and mechanical endurance behind the enhancing wound healing; Reduce scar proliferation, the control contracture has excellent biological compatibility; Can improve the artificial skin transplanting succeed rate, and improve healing quality; Surface structure and skin corium are entrenched togather, in conjunction with tight; Material source is extensive, production technique is simple; The treatment of the skin injury that can directly apply to prepared double-layered artificial skin inflammation, ulcer, burn and scald, reason such as iatrogenic cause is carried out personalized treatment to various disease; The shape size and the thickness of skin can prepare according to real needs.
Embodiment
Below in conjunction with instance technical scheme of the present invention is done further detailed description.
Embodiment 1:
Step 1). fibroblastic obtaining: get prepuce tissues after newborn infant's surgical blanking, subcutaneous lipids and muscle layer are removed with phosphate buffered saline buffer (PBS) flushing in the sterilization back, and skin is cut into strip, digest 2 hours with 4U/ml neutral protease liquid; Remove epidermal area, dermal tissue is shredded, with the collagenase liquid digestion of 625U/ml 2.5 hours, the cultivation of results inoblast; Former inoblast of being commissioned to train foster is used cell factory or method enlarged culturing such as revolving bottle culture systems or bio-reactor.
Step 2). take off the preparation of cell submucous layer of small intestine: the fresh pig jejunum is clean with distilled water flushing, be cut into required fragment, place and contained 0.2% Peracetic Acid and 10% alcoholic acid aqueous solution soaking disinfection 1 hour, with the rinsing of PBS liquid; Mechanical curettage mucous membrane of small intestine, flesh layer and serous coat, clean with the PBS rinsing, obtain submucosa; Again it is inserted in the NaOH solution of 1M, under 4 ± 2 ℃ of conditions, soaked 10 minutes, neutral with the rinsing of PBS liquid to pH; It is inserted in the mixing solutions that contains 40U/mlDNA enzyme and 10U/ml alpha-galactosidase soaked 1 hour, clean with the rinsing of PBS liquid; The cell submucous layer of small intestine is taken off in acquisition; Use void formers taking off evenly drilling on the cell submucous layer of small intestine; The aperture is 0.2mm; Spacing 1mm; Take off the cell submucous layer of small intestine of cell submucous layer of small intestine and atresia with foraminous and respectively get identical size stack, compress it is combined closely, dry back ethylene oxide sterilizing forms bilayer and takes off the cell submucous layer of small intestine;
Step 3). the preparation of bilayer skin nutrient solution; Each component by volume per-cent includes: commercial minimum essential nutrient solution DMEM 67.5%, commercial nutrient solution F12 22.5%, foetal calf serum 10%, Prostatropin 10ng/ml, epithelical cell growth factor 5ng/ml, Regular Insulin 5ng/ml, HYDROCORTISONE INJECTIONS 20ng/ml, VITAMIN B4 0.25mM, Transferrins,iron complexes 5 μ g/ml;
Step 4). the structure of double-layered artificial skin: under 4 ℃ of conditions; Press mass ratio with collagen 10: chitosan 1: mucinase 2: CHS 0.5 mixes, and uses the acetic acid of 0.1M that it is mixed with the solution of concentration as 10mg/ml, uviolizing under the ice bath; Foetal calf serum by its volume adding 10%; Adding final concentration again is the DMEM substratum of 10mg/ml, and regulating the pH value is 7.2～7.4, becomes gelating soln; The bilayer of preparation is taken off the cell submucous layer of small intestine and soaks into gelating soln, Procuring 30 minutes under 37 ℃ of environment in petridish; The inoblast of vitro culture is pressed 10
6The concentration of individual/ml is mixed in the above-mentioned gelating soln, presses 1ml/cm again
2The bilayer that is added drop-wise to Procuring takes off on porose of cell submucous layer of small intestine, in 5%CO
2Solidify after 30 minutes under 37 ℃ of conditions in the environment, insert the bilayer skin nutrient solution and cultivate after 3 days, be placed on again and continue on the cultivation support of cultivating in the vessel to cultivate 5 days, during every day change liquid, double-layered artificial skin is cultivated and is accomplished; Above-mentioned culture condition is 37 ℃, 5%CO
2Environment.
The double-layered artificial skin of this examples preparation comprise the human fibroblasts and synthesize, excretory extracellular matrix and growth factor, be convenient to clinical use, can participate in the reparation of the surface of a wound directly, be suitable for the damaged repair materials of skin histology.
Embodiment 2:
Step 1). fibroblastic obtaining: reference literature Dermal matrix as a carrier forin vivo delivery of human adipose-derived stem cells.Biomaterials.2008Apr; 29 (10): 1431-42. has proved that fat mesenchymal stem cell can be divided into inoblast.Get in the operation the healthy fatty tissue of depleted, the sterilization back shreds fatty tissue with phosphate buffered saline buffer (PBS) flushing, with the collagenase liquid digestion of 625U/ml 150 minutes, the fat mesenchymal stem cell of gathering in the crops is seeded to the culturing bottle cultivation; Carry out enlarged culturing with the method for cell factory or revolving bottle culture systems or bio-reactor again.
Step 2). the acellular dermal preparation: fresh pig skin is clean with distilled water flushing, and machinery is removed epidermal area and hypodermis layer, is cut into 0.4mm thickness, 5cm
2Size places and contained 0.2% (v/v) Peracetic Acid and 10% (v/v) alcoholic acid aqueous solution soaking disinfection 1 hour, uses the PBS rinsing; Place-80 ℃ of freezing half a hour again, guarantee that the material internal and external temperature reaches consistent, take out the back and thaws naturally, multigelation like this 3 times makes the cell disintegration of breaking fully, cleans with PBS solution; It is inserted in the NaOH solution of 0.3M, under 4 ± 2 ℃ of conditions, soaked 30 minutes, neutral with the PBS rinsing to pH; Again it is inserted in the solution that contains 30U/ml DNA enzyme and 20U/ml alpha-galactosidase and soaked 1 hour, clean with the PBS rinsing, obtain acellular dermal; Use void formers evenly drilling on acellular dermal; The aperture is 0.5mm, spacing 3mm, with the acellular dermal and the foraminous acellular dermal of atresia respectively get identical size stack, compress it combined closely; Dry back ethylene oxide sterilizing forms double-deck acellular dermal material;
Step 3). the preparation of bilayer skin nutrient solution; Each component by volume per-cent includes: commercial minimum essential nutrient solution DMEM67.5%, commercial nutrient solution F1222.5%, foetal calf serum 10%, Prostatropin 10ng/ml, epithelical cell growth factor 10ng/ml, Regular Insulin 5ng/ml, HYDROCORTISONE INJECTIONS 20ng/ml, VITAMIN B4 0.15mM, Transferrins,iron complexes 5 μ g/ml;
Step 4). the structure of double-layered artificial skin: under 4 ℃ of conditions; Press mass ratio with collagen 8: chitosan 1: mucinase 3: CHS 1 mixes, and uses the acetic acid of 0.3M that it is mixed with concentration and is 6mg/ml solution, uviolizing under the ice bath; Foetal calf serum by its volume adding 10%; Add the DMEM substratum again and make its final concentration reach 10mg/ml, regulating the pH value is 7.2～7.4, becomes gelating soln; With the preparation double-deck acellular dermal soak into gelating soln after, Procuring under 37 ℃ of environment; The fat mesenchymal stem cell of vitro culture is pressed 5 * 10
5The concentration of individual/ml is mixed in the gelating soln, again this gelating soln is pressed 1ml/cm
2Be added drop-wise on porose of double-deck acellular dermal of Procuring, place 5%CO
2After solidifying under 37 ℃ of conditions of environment, add the bilayer skin nutrient solution and cultivate after 2 days, be placed on again and continue on the cultivation support of cultivating in the vessel to cultivate 7 days, during every day change liquid, double-layered artificial skin is cultivated and is accomplished; Above-mentioned culture condition is 37 ℃, 5%CO
2Environment.
The double-layered artificial skin of this examples preparation includes people's fat mesenchymal stem cell; Can be divided into the healing that inoblast is participated in the surface of a wound directly at the surface of a wound; Have unrestricted, the molecular marker for increased proliferation in fat mesenchymal stem cell source simultaneously, promote the strong advantage of wound healing ability.
Claims (5)
1. double-layered artificial skin; Include inoblast; It is characterized in that; It is by taking off the membranaceous bio-derived material of cell as the top layer, is compound in the inner skin corium, both chimeric formations of forming of biologic bracket material by the extracellular matrix and the ESC of inoblast and synthesis secretion thereof; Described take off the membranaceous bio-derived material of cell be natural biological tissue take off cell submucous layer of small intestine, acellular dermal matrix through taking off the formed membranaceous bio-medical material of cell, comprising, take off the cell manadesma, take off the cell submucous layer of bladder, take off the cell amnion, take off cell dural any; Described inoblast comprises can be to the stem cell of inoblast differentiation; Described biologic bracket material is three-dimensional propping material to be provided for cell growth, comprises any or several kinds mixing of Fibrinogen, scleroproein, alginates, collagen protein, mucinase, CHS, chitosan, hydrogel, gelatin, POLYACTIC ACID, PGTA.
2. the method for preparing the described double-layered artificial skin of claim 1 includes the step that cell obtained, took off the preparation of cell biological derived material, it is characterized in that concrete steps are following:
Step 1). fibroblastic obtaining: the human fibroblasts carries out vitro culture and extensive amplification by the cultural method of routine;
Step 2). take off the preparation of cell biological derived material: adopt dermal layer of the skin, manadesma, submucous layer of small intestine, amnion, dural any membranaceous biomaterial preparation; Material placed contain Peracetic Acid and alcoholic acid aqueous solution soaking disinfection was not less than 1 hour, after cleaning with phosphate buffered saline buffer; Being placed on the NaOH solution soaking handles; Embathe to pH neutral with phosphate buffered saline buffer; It being inserted in the mixing solutions that contains DNA enzyme and alpha-galactosidase handles more than 25 minutes again; Evenly drilling above that after embathing with phosphate buffered saline buffer; With the taking off cell biological derived material stack with foraminous and compress it is combined closely of atresia, become bilayer behind the dry sterilization and take off the cell biological derived material again;
Step 3). the preparation of double-layered artificial skin nutrient solution; Its component by volume per-cent includes: commercial minimum must nutrient solution DMEM 67.5%, commercial nutrient solution F12 is 22.5%, foetal calf serum 5～15%, Prostatropin 2～100ng/ml, epithelical cell growth factor 2～100ng/ml, Regular Insulin 1～50ng/ml, HYDROCORTISONE INJECTIONS 10～500ng/ml, VITAMIN B4 0.2～0.25mM and Transferrins,iron complexes 1～10 μ g/ml;
Step 4). the structure of double-layered artificial skin: the gelating soln of preparing biologic bracket material earlier; After the bilayer of preparation taken off the cell biological derived material and soak into gelating soln, place Procuring under 37 ℃ of environment; The inoblast of vitro culture is pressed 10
4～10
6The concentration of individual/ml is mixed in the gelating soln, again by 0.1～1ml/cm
2Be added drop-wise on porose of Procuring material, place 5%CO
2Solidify under 37 ℃ of conditions in the environment; Insert again cultivate 2～3 days in the double-layered artificial skin nutrient solution after, be placed on and continue on the cultivation support of cultivating in the vessel to cultivate 5～8 days, change liquid every day between during cultivation, cultivates and accomplish; Above-mentioned culture condition is 37 ℃, 5%CO
2Environment.
3. preparation method according to claim 2 is characterized in that step 2) in be 0.1～1mm in the said aperture of taking off the drilling of cell biological derived material, spacing is 0.5～5mm.
4. preparation method according to claim 2 is characterized in that, in step 2) described in Peracetic Acid and alcoholic acid volume final concentration be respectively 0.1～0.5% and 2～10%; The final concentration of said DNA enzyme and alpha-galactosidase is respectively 30～50U/ml and 10～25U/ml.
5. preparation method according to claim 2; It is characterized in that, in step 2) described in soaking disinfection clean after, before the NaOH solution soaking; Material is placed-80 ℃ more than freezing half a hour; Take out the back and thaws naturally, so multigelation is 2～5 times, with after the phosphate buffered saline buffer cleaning again in the NaOH solution soaking.
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| CN114836378B (en) * | 2022-04-11 | 2023-07-04 | 郑州源创基因科技有限公司 | In-vitro culture method of autologous breast milk stem cells, injection and application of injection in skin injury repair |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN1468634A (en) * | 2002-07-15 | 2004-01-21 | 上海组织工程研究与开发中心 | Double-layered artificial skin and its prepn process |
| CN1607012A (en) * | 2003-10-13 | 2005-04-20 | 刘凯 | Method for preparing human body tissue engineering skin |
| CN1868548A (en) * | 2006-05-30 | 2006-11-29 | 中国人民解放军第二军医大学 | Heterologous or heterogenic decelled epidermis substitute used for human fibroblast-like cell modification |
-
2008
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| CN1468634A (en) * | 2002-07-15 | 2004-01-21 | 上海组织工程研究与开发中心 | Double-layered artificial skin and its prepn process |
| CN1607012A (en) * | 2003-10-13 | 2005-04-20 | 刘凯 | Method for preparing human body tissue engineering skin |
| CN1868548A (en) * | 2006-05-30 | 2006-11-29 | 中国人民解放军第二军医大学 | Heterologous or heterogenic decelled epidermis substitute used for human fibroblast-like cell modification |
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