CN107475183A - A kind of fat survival and regeneration method for improving adipose tissue - Google Patents

A kind of fat survival and regeneration method for improving adipose tissue Download PDF

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Publication number
CN107475183A
CN107475183A CN201710904761.3A CN201710904761A CN107475183A CN 107475183 A CN107475183 A CN 107475183A CN 201710904761 A CN201710904761 A CN 201710904761A CN 107475183 A CN107475183 A CN 107475183A
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adipocyte
fat
adipose tissue
transplanting
cell
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张继伟
王红娟
李雪梅
王占利
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Shijiazhuang Reibel Medical Beauty Clinic
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Shijiazhuang Reibel Medical Beauty Clinic
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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
    • A61L27/3616—Blood, e.g. platelet-rich plasma
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
    • A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3895—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
    • 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
    • C12N5/0653—Adipocytes; Adipose tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/06—Flowable or injectable implant compositions
    • 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
    • A61L2430/04—Materials or treatment for tissue regeneration for mammary reconstruction

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Abstract

The invention discloses a kind of fat survival and regeneration method for improving adipose tissue, and it includes extraction PECTORAL LIMB SKELETON, adipocyte transplanting, extraction nutritional ingredient and backfill transplanting.The adipocyte that the inventive method obtains, the cell of fat can be constantly divided into after transplanting, the transplanting for solving conventional fat absorbs activity and adipocyte transplants the problems such as absorbing or be downright bad；Long-term, lasting, the stable survival of the fat of transplanting can be made.Adipocyte is converted into after the material transplanting that the present invention extracts, original adipocyte continued growth can be promoted, final stabilization turns into adipose tissue, and survival rate is up to more than 90%.Activated it is adipocyte transforming be adipocyte, the adipose tissue at position need to be filled by being formed, promote original adipocyte continued growth, it can help to form blood vessel, accelerate the vascularization of transplant fat and survive again, it is releasable to promote angiogenesis factor, it can make the fat of transplanting is quick to survive as normal, vital adipose tissue, patient's body aesthetic feeling can be obviously improved.

Description

A kind of fat survival and regeneration method for improving adipose tissue
Technical field
Survived the present invention relates to adipocyte autotransplantation and regeneration method.
Background technology
After first reported Free fat success in 1893 from Neuber, numerous doctor U.S. experts uses this method Carry out the filling reparation at the positions such as substantial amounts of chest, face, but all not up to expected surgical effect.Fat is mainly distributed on people The positions such as body hypodermis, omentum majus, mesenterium and kidney week.Adipose tissue is by PECTORAL LIMB SKELETON and mature fat cell structure Into, containing 90% fat drips in the bag slurry in mature fat cell, its transplant after can rapid metabolization fall.Mature fat cell is point Change thesocyte, lose the ability for continuing propagation.Mature fat cell can only increase the volume of transplantation site and add Upper mature fat cell is easily damaged by external mechanical, and causes significant fraction mature fat cell to sustain damage and destroy.Often Fat transfer is treated lipochondrion, removes the impurity such as grease or watery blood, and meeting partial necrosis, is survived after transplanting Rate is 30%~70% or so.This is also the volume of transplantation site occur after explaining the simple adipose tissue transplantation filling of tradition in the past Diminish, fatty fibrosis the problems such as.
The content of the invention
The present invention provide a kind of fat survival for making PECTORAL LIMB SKELETON to improve adipose tissue under certain condition and Regeneration method, to solve the problems, such as that prior art is present.
To realize the object of the invention, this fat survival for improving adipose tissue and regeneration method include following step Suddenly：
A. PECTORAL LIMB SKELETON is extracted
40 minutes aseptically will be stood in 2~5 DEG C of sterile salines in clean adipose tissue mixed liquor After centrifuge, centrifuge 10 minutes after collect PECTORAL LIMB SKELETON；
B. adipocyte is transplanted
After the above-mentioned PECTORAL LIMB SKELETON isolated is mixed with cleaning solution by 1: 1 volume ratio, stood at 37~39 DEG C 30~60 minutes, after cell confluency reaches 70%~80%, it is transplanted on hyaluronic acid；
C. nutritional ingredient is extracted
After the blood samples of patients of extraction is centrifuged 10~15 minutes, then lower floor's heavy component extracted at 40~60 DEG C, Obtain glucocorticoid, growth hormone, prostaglandin, insulin, thyroxine, IGF and fibroblast The mixture of the factor；
D. backfill migrates to filling position
Said mixture backfill is transplanted into patient itself need to fill position.
Preceding cell is PECTORAL LIMB SKELETON, or fat mesenchymal stem cell (Mesenchymal adipose cell Precursors ADSCs) it is that Hausberger proposes in nineteen fifty-five.Preceding cell volume is small, and differentiation degree is low, to wound and lacks The tolerance of oxygen is better than mature fat cell.Preceding cell is a kind of multipotent stem cells in adipose tissue.Because adipose tissue comes Source is extensively, materials are easy, is not related to ethics problem, is easy to autotransplantation, and preceding cell has certainly as other adult stem cells I updates, vigor is lasting and multi-lineage potential.Largely it is demonstrated experimentally that under suitable inductive condition, preceding cell is respectively to 3 The cell differentiation of individual germinal layer, as APSC pluripotent cells, epithelial cell, smooth muscle cell, cardiac muscle cell, cartilage cell, skeletonization are thin Born of the same parents, adipocyte, nerve cell, vascular endothelial cell and liver cell etc..
The technological progress that the present invention obtains：The adipocyte that the inventive method obtains, be have perfect blood circulation and Nutrition supply function, the cell of fat can be constantly divided into after transplanting, the transplanting for solving conventional fat absorbs activity fat carefully Born of the same parents transplant the problems such as absorbing or be downright bad, can make long-term, lasting, the stable survival of the fat of transplanting.The material that the present invention extracts moves Adipocyte is converted into after plant, original adipocyte continued growth can be promoted, final stabilization turns into adipose tissue, and survival rate is high Up to more than 90%.Activated it is adipocyte transforming be adipocyte, the adipose tissue at position need to be filled by being formed, and be promoted original It adipocyte continued growth, can help to form blood vessel, accelerate the vascularization of transplant fat and survive again, it is releasable to promote blood vessel shape Into the factor, it can make the fat of transplanting is quick to survive as normal, vital adipose tissue, patient's pretty figure can be obviously improved Sense, it is a kind of method for the lifting patient's pretty figure for being significantly better than traditional simple adipose tissue transplantation process.
Embodiment
This fat survival for improving adipose tissue and regeneration method comprise the following steps：
A. PECTORAL LIMB SKELETON is extracted
Aseptically extracted from human fatty tissue in postoperative adipose tissue mixed liquor by cleaning solution (d- Hank ' s cleaning solutions, formed for the configuration such as sodium chloride and disodium bicarbonate and potassium chloride) arcotic and haemocyte are washed and done After net, 40 minutes are stood in 2~5 DEG C of sterile salines；Then reuse centrifuge and 1500 turns/min, centrifugation ten are set After minute, reject supernatant, PECTORAL LIMB SKELETON is obtained.
Extracted from human fatty tissue and PECTORAL LIMB SKELETON is isolated in postoperative adipose tissue mixed liquor, adipose tissue tool Have a convenient material drawing and the high advantage of stem cell content, from the point of view of average experiment data, can be obtained per 30ml adipose tissues 2X108~ 6X108 fat mesenchymal stem cell.Adipose-derived mescenchymal stem cell content is higher than the mescenchymal stem cell of derived from bone marrow. And donor age size does not have much affect to adipose-derived mescenchymal stem cell content.PECTORAL LIMB SKELETON is not easily susceptible to outer Boundary's mechanical damage and with materials easily, amount is big, can draw materials repeatedly, it is impaired it is relatively light, can cell propagation and carry out autotransplantation without The advantages that immunological rejection.
Experimental technique and principle：With cell culture technology, the flow cytometry (phenotype of PECTORAL LIMB SKELETON after amplification in vitro Can change, be mainly reflected in the change of cell surface protein and cytokine-expressing), differential centrifugation art (can be by matrix blood Solencyte precipitates to be separated with the mature fat cell of suspension, in precipitation in addition to PECTORAL LIMB SKELETON, in addition to haemocyte, into fiber finer Born of the same parents and endothelial cell, stromal vascular cell precipitation can be inoculated into plastic culture bottle, and stroma cell can be adherent, hematopoiesis and other Contaminating cell is not adherent, is removed in subsequent succeeding generations, and the PECTORAL LIMB SKELETON finally given can be protected within some time Hold film differentiation state).The PECTORAL LIMB SKELETON being separately cultured out in adipose tissue mixed liquor by Induction of committed differentiation technology, its The condition for improving transplanting success is need to adsorb that cell migration could be carried out on the support of inoculation, second, must be built for it rapidly Nutriment necessary to vertical blood supply, third, quickly accelerating PECTORAL LIMB SKELETON to turn into mature fat cell, and reach propagation point The purpose of change.
B. adipocyte is transplanted
By the above-mentioned PECTORAL LIMB SKELETON isolated with a sterile centrifugation tube, the cleaning solution (cleaning solution loaded after dilution Be formulated for sodium chloride and disodium bicarbonate and potassium chloride etc.) adjustment concentration be 120mg/ml and count；It will centrifuge The PECTORAL LIMB SKELETON gone out is mixed with 1: 1 ratio.At 37~39 DEG C, after cell fusion reaches 70%~80%, training is used Foster PECTORAL LIMB SKELETON is inoculated into 6 orifice plates by 1 × 105 cells/well, is transplanted to the biomaterial hyaluronic acid as support On；
The above-mentioned PECTORAL LIMB SKELETON isolated is transplanted on the biomaterial hyaluronic acid as support, searching can provide The biomaterial of stent migration carries out adipocyte transplanting, and PECTORAL LIMB SKELETON, which is only adsorbed on appropriate support, could be carried out carefully Born of the same parents migrate.At present, being used as the biomedical Biodegradable material of fat tissue cell's support mainly has by popular institute The collagen of receiving, hyaluronic acid etc..(the present embodiment is only to hyaluronic acid as the medical biotechnology material conduct for providing support Research object illustrates) because hyaluronic acid can provide substantial amounts of hole, be advantageous to PECTORAL LIMB SKELETON connection and it is fixed and Into a kind of polymer, due to including the medical drugs materials such as the extraordinary coupling agent of histocompatbility in hyaluronic acid, and make inflammatory Reaction drops to very low level.
C. nutritional ingredient is extracted
After the human blood of extraction is centrifuged 10~15 minutes with 3000 revs/min, then by lower floor's heavy component at 40~60 DEG C After extraction, obtain glucocorticoid, growth hormone, prostaglandin, insulin, thyroxine, IGF and into The mixture of the fibrocyte factor etc.；
The adipose tissue of body is the tissue of very vascular, and the density of blood supply therein and capillary is to determine The key of the survival rate of adipose tissue.It is reported that each adipocyte is at least connected with a capillary, this is also card The basic reason that the bright simple fat transfer effect of tradition in the past is poor, survival rate is low.Blood separation elite extraction, by the patient of extraction Blood is centrifuged 10~15 minutes on centrifuge with 3000 revs/min, and its upper strata is rich in amino acid, vitamin, PRP, lipid material Deng being the necessary material of Hemapoiesis.Its lower floor obtains glucocorticoid, growth hormone, prostaglandin, pancreas by extracting again Island element, thyroxine, IGF and fibroblast factor etc., the existing direct regulation and control of PECTORAL LIMB SKELETON can be promoted Effect, nutritional ingredient is directly drawn, and various growth factors are provided, promote cellular vascular quickly to be formed, it is normal so as to be formed Adipose tissue and the lasting form and volume that maintain transplantation site, its various factor accelerate mature fat cell propagation again Volume becomes the big nutriment condition for providing basis.
D. backfill migrates to filling position
The i.e. deployed idiotrophic material of the mixture obtained after above-mentioned extraction is backfilled by medical science invasive methods and moved Position need to be filled by planting patient itself.

Claims (1)

1. a kind of fat survival and regeneration method for improving adipose tissue, it is characterised in that it comprises the following steps：
A. PECTORAL LIMB SKELETON is extracted
After standing 40 minutes in 2~5 DEG C of sterile salines in adipose tissue mixed liquor that aseptically will be clean from The heart is separated, and PECTORAL LIMB SKELETON is collected after centrifuging 10 minutes；
B. adipocyte is transplanted
After the above-mentioned PECTORAL LIMB SKELETON isolated is mixed with cleaning solution by 1: 1 volume ratio, at 37~39 DEG C stand 30~ 60 minutes, after cell confluency reaches 70%~80%, it is transplanted on hyaluronic acid；
C. nutritional ingredient is extracted
After the blood samples of patients of extraction is centrifuged 10~15 minutes, then lower floor's heavy component extracted at 40~60 DEG C, obtained Glucocorticoid, growth hormone, prostaglandin, insulin, thyroxine, IGF and the fibroblast factor Mixture；
D. backfill migrates to filling position
Said mixture backfill is transplanted into patient itself need to fill position.
CN201710904761.3A 2017-09-20 2017-09-20 A kind of fat survival and regeneration method for improving adipose tissue Pending CN107475183A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112773937A (en) * 2021-01-26 2021-05-11 北京臻悦医疗美容诊所有限公司 Method for improving survival rate of injected fat

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112773937A (en) * 2021-01-26 2021-05-11 北京臻悦医疗美容诊所有限公司 Method for improving survival rate of injected fat

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Application publication date: 20171215