CN106237391A - Adipose tissue composite preparation and preparation method and application thereof - Google Patents
Adipose tissue composite preparation and preparation method and application thereof Download PDFInfo
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- CN106237391A CN106237391A CN201610614517.9A CN201610614517A CN106237391A CN 106237391 A CN106237391 A CN 106237391A CN 201610614517 A CN201610614517 A CN 201610614517A CN 106237391 A CN106237391 A CN 106237391A
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Abstract
The invention relates to an adipose tissue composite preparation, a preparation method and application thereof. The adipose tissue composite preparation comprises PDGF, umbilical cord mesenchymal stem cells, SVF and adipose tissue. SVF, umbilical cord mesenchymal stem cells and PDGF are prepared into cell suspension, and then adipose tissues and the cell suspension are uniformly mixed according to the volume ratio of 20:1 to prepare the adipose tissue coincidence preparation. The adipose tissue composite preparation is applied as a filling preparation in the plastic or cosmetic field. The composite biological preparation has the advantages of easy revascularization after injection, high survival rate, safety and reliability, and the fat tissue is light yellow, soft and glossy after being transplanted for 2 to 3 months and is only wrapped by a few fibers; the density of the microvascular at the central part of the transplanted fat is normal, and the revascularization is obvious.
Description
Technical field
The present invention relates to field of biological, particularly relate to a kind of fatty tissue compound formulation and preparation method thereof and answer
With.
Background technology
Autologous fat transplantation is clinical commonly used a kind of fat transplantation scheme, and the advantage of this method is bio-compatible
Property good, tissue-derived abundant, it is easy to moulding, the untoward reaction such as immunologic rejection will not be produced；Shortcoming is to be injected into face or breast
The lipochondrion at position can not well set up blood supply system.When significant quantities of fat is stacked into together, can because of blood supply not
Foot causes adipose cell liquefaction and necrosis, dissolves, absorbs, and easily causes the sequela such as infection, deformation, pain, often needs again to note
Penetrate and maintain shape.
The Chinese invention patent application of Application No. 201110203210.7 discloses a kind of fat being enriched with autologous stem cells
Fat tissue is enlarged the bosom the preparation method of material, this patent is separated by a small amount of liposuction first respectively and amplification culture fat stem cell,
Before transplanting, vascular stroma composition (stromal vascular fraction, SVF) and lipochondrion are prepared in liposuction respectively, then will
All cells mixes with lipochondrion, prepares material of enlarging the bosom.But this invention needs to implement twice liposuction, and add thin
The lazy weight of born of the same parents, cytoactive are relatively low, and the cell of re-injection is difficult to proliferation and differentiation and forms new tissue, do not reach and fill improvement
Effect.
After pure grain fatty tissue injection transplantation, fatty tissue can only lean on the infiltration of surrounding tissue liquid and infiltration to maintain
Nutrition supply, and the distance of this supply only has 150～200 μm, exceedes this distance and is accomplished by generating new blood vessel to provide battalion
Support.But, neovascularization growth is slow, and every day only grows tens microns, causes transplant fat peripheral part blood supply to rebuild slowly, blood
For postponing, transplant fat center is chronically at ischemic state.The fatty tissue of transplant fat core due to long-time ischemia,
Anoxia, there occurs necrosis, liquefaction, is then removed by macrophage, occurs fibrocystic to become, and the appearance of microcapsule imply that transplant
Final fibrosis and being absorbed.Common recognition has gradually been reached in increasing research, transplant set up in early days fully with blood supply timely,
Graft revascularization in time is to affect its key surviving volume, is also to reduce the key that its fibrocystic becomes.
Summary of the invention
In view of this, it is necessary to for above-mentioned problem, it is provided that a kind of fatty tissue compound formulation and preparation method thereof and
Application.
In order to realize above-mentioned purpose, the present invention is achieved through the following technical solutions:
A kind of fatty tissue compound formulation, comprises platelet derived growth factor (Platelet derived growth
Factor, PDGF), umbilical cord mesenchymal stem cells (Mesnchymal Stem Cells, MSCs) and stromal vascular component
(stromal vascular fraction, SVF).
Preferably, described fatty tissue compound formulation comprises platelet derived growth factor, umbilical cord mesenchymal stem cells, base
Matter vascular component and fatty tissue.
It is further preferred that the content of platelet derived growth factor is 5-20ng/ in described fatty tissue compound formulation
ml。
It is further preferred that the quantity of umbilical cord mesenchymal stem cells is 5 × 10 in described fatty tissue compound formulation5～5
×106Individual/ml.
As preferably, the content of fatty tissue compound formulation mesostroma vascular component of the present invention is 5 × 105～5 ×
106Individual/ml.
Preferably, described fatty tissue compound formulation contains the following component of following content:
Umbilical cord mesenchymal stem cells 1 × 106～5 × 106Individual/ml,
Stromal vascular component 1 × 106Individual/ml,
Platelet derived growth factor 5～20ng/ml.
Preferably, described fatty tissue compound formulation contains the following component of following content:
Umbilical cord mesenchymal stem cells 1 × 106Individual/ml,
Stromal vascular component 1 × 106Individual/ml,
Platelet derived growth factor 10ng/ml.
It is further preferred that described stromal vascular component extracts from fatty tissue.
The preparation method of above-mentioned fatty tissue compound formulation, comprises the following steps: by stromal vascular component, umbilical cord mesenchyma
Stem cell and platelet derived growth factor are configured to cell suspension, then by fatty tissue with cell suspension according to volume ratio 20:1
Ratio mixing.
Above-mentioned fatty tissue compound formulation shaping or beauty treatment in as filling preparation application.
Described shaping or beauty treatment include the depressed deformity of filling facial area, mamaplasty, rich temporo and augmentation rhinoplasty.
Platelet derived growth factor is a kind of important factor,mitogenic, has stimulation specific cells group and divides increasing
The ability grown, can promote the generation of fibroblast, and can promote that subcutaneous capillary is formed, and repairs that subcutaneous blood is micro-follows
Loop systems.
Umbilical cord mesenchymal stem cells is isolatable from umbilical cord tissue, its wide material sources, and multiplication capacity is strong, and immunogenicity is weak, and
Drawing materials conveniently, there is not immunologic rejection, and has immunoregulation effect in heteroplastic transplantation, can improve the one-tenth that cell or tissue is transplanted
Power.Additionally, umbilical cord mesenchymal stem cells can be differentiated to form new tissue after re-injection, the various active factor of secretion, can promote
Enter vascularization and tissue repair.
Stromal vascular component be an adipose-derived stem cells with vascular endothelial cell, peripheral blood cells, T cell and other
The colony that inflammatory cell mixes mutually, can be divided into mature fat cell in vivo under environment, promote that host stem cells is to by district
Directional migration, subparticipation composition neovascular endothelium cell, contribute to the foundation of transplant fat tissue blood supply in early days.
The allosome umbilical cord mesenchymal stem cells of infusion in compound formulation of the present invention, cell concentration is relatively big, and multiplication capacity is strong, can lure
Lead and be divided into fatty tissue, new tissue, and the various active factor secreted can be differentiated to form after re-injection, there is immunomodulating
Effect, promotes vascularization and tissue repair.It is multiple that the platelet derived growth factor added and mescenchymal stem cell are secreted
Cytokine, forms good microenvironment, and the fat stem cell in SVF and Derived from Mesenchymal Stem Cells can be promoted for ripe fat
Fat cell, plays tissue substitute effect.Mescenchymal stem cell can suppress inflammatory reaction simultaneously, promotes subcutaneous new capillary vessel
Formed, make the fatty tissue after transplanting rebuild rapidly blood supply, improve the survival rate of transplant fat, it is to avoid tissue liquefaction and necrosis.
Compared with prior art, there is advantages that
(1) present invention provides a kind of containing SVF, umbilical cord mesenchymal stem cells and platelet derived growth factor auxiliary in fat transplantation
Composite biological agent in tissue filling.The advantage that this compound formulation is prone to revascularization after having injection, survival rate improves, peace
Complete reliable, it is adaptable to Soft-tissue operation, to repair depressed deformity, face and health that soft tissue defects causes and improve looks moulding, as can
For the depressed deformity etc. of filling facial area, it is also used for mamaplasty, rich temporo and augmentation rhinoplasty etc., can promote that profile is extensive in beauty treatment filling
Again, liquefaction and necrosis is prevented.
(2) compound formulation of the present invention survival rate in human body is high.By laboratory observation, there is after transplanting a large amount of normal-fat
Cell, adipose cell is clear, the most a small amount of bad color, inflammatory reaction phenomenon and fibre gap；After transplanting 2-3 month, fatty tissue
Color is yellowish, soft, glossy, only a few fibres parcel；After fatty tissue preparation re-injection of the present invention is filled, transplant fat center
The microvessel density at position is normal, and revascularizationization is obvious.The manufacture method of fatty tissue preparation of the present invention is simple, can be a large amount of
Preparation.
Accompanying drawing explanation
Fig. 1 is matched group FSC-SSC figure in embodiment 1 flow cytometer detection.
Fig. 2 is the flow cytometer detection result figure of matched group CD73, CD45 in embodiment 1 flow cytometer detection.
Fig. 3 is the flow cytometer detection result figure of matched group CD19, CD11b in embodiment 1 flow cytometer detection.
Fig. 4 is the flow cytometer detection result figure of matched group CD34, CD90 in embodiment 1 flow cytometer detection.
Fig. 5 is the flow cytometer detection result figure of matched group CD105, HLA-DR in embodiment 1 flow cytometer detection.
Fig. 6 is experimental group FSC-SSC figure in embodiment 1 flow cytometer detection.
Fig. 7 is the flow cytometer detection result figure of experimental group CD73, CD45 in embodiment 1 flow cytometer detection.
Fig. 8 is the flow cytometer detection result figure of experimental group CD19, CD11b in embodiment 1 flow cytometer detection.
Fig. 9 is the flow cytometer detection result figure of experimental group CD34, CD90 in embodiment 1 flow cytometer detection.
Figure 10 is the flow cytometer detection result figure of experimental group CD105, HLA-DR in embodiment 1 flow cytometer detection.
Figure 11 be in embodiment 1 umbilical cord mesenchymal stem cells adipogenic induction differentiation in matched group dyeing picture.Wherein, figure
11A, Figure 11 B and Figure 11 C be respectively 100 ×, 200 ×, the 400 × lower picture observed.
Figure 12 be in embodiment 1 umbilical cord mesenchymal stem cells adipogenic induction differentiation in experimental group dyeing picture.Wherein, figure
12A, Figure 12 B and Figure 12 C be respectively 100 ×, 200 ×, the 400 × lower picture observed.
Detailed description of the invention
In order to better illustrate the present invention, it is described further with detailed description of the invention below in conjunction with the accompanying drawings.
In the present invention, source or the preparation method of each component are as follows:
(1)PDGF
Selecting MCSD002 vectors containing human platelet-derived growth, manufacturer is wheat bin bio tech ltd, Shanghai,
0.5mg/ props up, and preserves: 2-8 DEG C.
(2) umbilical cord mesenchymal stem cells
In the present embodiment the separation and Extraction of umbilical cord mesenchymal stem cells and Secondary Culture with reference to Sun Guodong etc. " between people's umbilical cord
The separation and Culture of mesenchymal stem cells and become, to skeletonization, the experimentation that fat breaks up " (Sun Guodong, Li Zhizhong, Wang Jing, etc. people's umbilical cord
The separation and Culture of mescenchymal stem cell and become the experimentation [J] that fat breaks up to skeletonization. XI AN JIAOTONG UNIVERSITY Subject Index: medicine,
2010,31 (2), 143-147) in report related content to obtain umbilical cord mesenchymal stem cells carry out surface antigen detection and
Adipogenic induction breaks up.
1. umbilical cord mesenchymal stem cells surface antigen is identified
The concrete grammar of umbilical cord mesenchymal stem cells surface antigen detection is as follows: takes P3 for umbilical cord stem cells, sucks cultivation
Base, with 0.25% trypsin solution conventional digestion, makes 1 × 105Cell suspension, take respectively anti-human CD73, CD45, CD19,
The each 5 μ L of monoclonal antibody of CD34, CD90, CD11b, CD105 and HLA-DR, add cell suspension 500 μ L, lucifuge under room temperature
Hatching 20min, set up blank Isotype control simultaneously, 1500r/min is centrifuged 5min, abandons supernatant, washs 2 with the PBS containing 10%FBS
Time, with the resuspended rear upper machine testing of 500 μ L PBS.P3 is for umbilical cord mesenchymal stem cells flow cytometer detection result as Figure 1-10 shows.Root
According to 2006, the mescenchymal stem cell that international artery cell association formulates identified that minimum standards is: CD73, CD90, CD105 are more than
In 95%, CD45, CD19, CD34, CD11b and HLA-DR are less than or equal to 2%.
From Fig. 1-10 streaming result, umbilical cord mesenchymal stem cells CD73, CD90, CD105 of separation and Culture of the present invention
>=95.0%；CD45, CD19, CD34, CD11b and HLA-DR≤2.0%, point out the cell added in compound formulation not go out
Existing differentiating phenomenon, meets the feature of mescenchymal stem cell.
2. the one-tenth fat of umbilical cord mesenchymal stem cells is identified
The concrete grammar of the adipogenic induction differentiation of umbilical cord mesenchymal stem cells is as follows: take P3 for umbilical cord mesenchymal stem cells,
Digestion, with 106Individual cells/well density is inoculated in 6 well culture plates, and after cultivating 24h, cell attachment also stretches, and is replaced with fat and divides
Change culture medium (to be 10% hyclone, 10mg/L insulin containing DMEM-F12 culture medium, volume fraction, fill in rice to 1 μm ol/L
Pine, 100 μm ol/L indomethacins, 500 μm ol/L IBMX), the complete medium group containing 10% hyclone is right as feminine gender
According to.Every 3d changes 1 subculture, and directed differentiation i.e. can use external evoked point of oil red O stain qualitative observation after inducing 21d
The result changed.Dyeing picture is as is illustrated by figs. 11 and 12.Matched group result is negative, and test results is positive, illustrates to obtain
Umbilical cord mesenchymal stem cells can be broken up by adipogenic induction.
(3) fatty tissue and SVF
In the present embodiment, fatty tissue is drawn materials and the separation and Extraction of SVF " is promoting the optimal of fat transplantation with reference to Zhu Ming etc.
The experimentation of SVFs concentration " (Zhu Ming, Lu Feng, high Jian Hua, etc. promote the experimentation of the optimal SVFs concentration of fat transplantation
[J]. China's shaping surgery magazine, 2012,28 (4), 284-290) related content reported carries out.It is summarized as follows:
Isopyknic pre-cold saline (4 DEG C) is joined in fatty tissue, after covering bottle cap, repeatedly rocks fat group
Knit, stand to substantially layering；Discarding fatty tissue lower floor solution, this step is repeated once；Add isopyknic in fat tissue
Pre-cold saline, 700g is centrifuged 3min, sucks upper-layer fat oil and lower floor's liquid with pipet；The fat of reserved preparation final volume
Fat, as the adipose tissue transplantation of compound formulation, shreds reserved fat with shears；The fatty tissue shredded is placed in cold water
In bath and put to 4 DEG C of Refrigerator stores.
Other fat add the 0.5%I Collagenase Type in-solution digestion of 1/2 fat volume, move in 37 DEG C of constant temperature oscillators,
200R digests 15-30min.After having digested, 800g is centrifuged 10min；Abandon upper-layer fat oil and remaining liq；Add normal saline weight
Outstanding precipitation, with 100 μm cell strainer filtration cell suspensions after being centrifuged, recentrifuge precipitation is SVF.
Embodiment 1, the conditional filtering test of fatty tissue compound formulation
In the present embodiment, compound formulation is made up of fatty tissue, umbilical cord mesenchymal stem cells, SVF and PDGF.In order to determine
The optimized scope of every kind of amounts of components, carries out conditional filtering test for inspection target to compound formulation with weight in wet base.Fill between umbilical cord
Matter stem cell, SVF with PDGF normal saline become the cell suspension of suitable concn, by fatty tissue and cell suspension according to
The ratio mix homogeneously of volume ratio 20:1, makes the final concentration of each component in compound formulation of the present invention as shown in table 1.
A composition selection combination in table 1, compound formulation
| Group | Compound formulation forms |
| Matched group | Fatty tissue |
| Experimental group 1 | 5×105Individual/ml MSCs+1 × 106Individual/ml SVF+1ng/mL PDGF+ fatty tissue |
| Experimental group 2 | 1×106Individual/ml MSCs+1 × 106Individual/ml SVF+5ng/mL PDGF+ fatty tissue |
| Experimental group 3 | 2.5×106Individual/ml MSCs+1 × 106Individual/ml SVF+10ng/mL PDGF+ fatty tissue |
| Experimental group 4 | 5×106Individual/ml MSCs+1 × 106Individual/ml SVF+20ng/mL PDGF+ fatty tissue |
Choose female 4～6 weeks nude mices 100, be randomly divided into 5 groups, often group 20.With 2% pentobarbital sodium 50ms/ks abdomen
Chamber injecting anesthetic, injects every nude mice back skin with syringe by the most random for the compound formulation in each experimental group and matched group
Under, every nude mice injection 0.5ml (about 450mg).After fat grafting 3 months, outside graft tunicle, separate graft, completely
Taking out graft and it is carried out weight in wet base mensuration, investigate transplant fat tissue survival rate, result is as shown in table 2.
Table 2, graft weight in wet base statistical table
| Group | Weight in wet base |
| Matched group | 89.25±9.50mg |
| Experimental group 1 | 93.73±13.67mg |
| Experimental group 2 | 167.05±14.62mg* |
| Experimental group 3 | 189.55±12.74mg* |
| Experimental group 4 | 230.41±15.50mg* |
* represent that compared with matched group P < 0.05 has significant difference.
Statistic software SPSS 17.0 is used to carry out data process the data of table 2.Use the side of randomized block design data
Difference analysis, compares weight in wet base.The graft weight in wet base that matched group obtains is about 89.25mg, the graft weight in wet base of experimental group 1
Being about 93.73mg, experimental group 1, compared with matched group, does not has significant difference, P > 0.05.The graft weight in wet base of experimental group 2 is about
167.05mg, the graft weight in wet base about 189.55mg of experimental group 3, the graft weight in wet base of experimental group 4 are 230.41mg, three groups with
Matched group is compared, and is respectively provided with significant difference, P < 0.05.
In sum, the optimal concentration scope of the fat each composition of compound formulation is as follows: 1 × 106～5 × 106Individual/ml umbilical cord
Mescenchymal stem cell+1 × 106Individual/ml SVF+5～20ng/mL PDGF
Embodiment 2, fatty tissue compound formulation and compliance test result
In the present embodiment, compound formulation is made up of fatty tissue, umbilical cord mesenchymal stem cells, SVF and PDGF.In order to enter one
The effect of the compound formulation of step card optimization formula, becomes to close by umbilical cord mesenchymal stem cells, SVF with PDGF normal saline
The cell suspension of suitable concentration, by fatty tissue and cell suspension according to the ratio mix homogeneously of volume ratio 20:1, makes the present invention multiple
Close the final concentration of each component in preparation as shown in table 3.
Each component combined effect checking packet in table 3, compound formulation
Choose female 4～6 weeks naked 120, be randomly divided into 6 groups, often group 20.With 2% pentobarbital sodium 50ms/ks abdominal cavity
Injecting anesthetic, injects every nude mice dorsal sc with syringe by the most random for the compound formulation in each experimental group and matched group,
Every nude mice injection 0.5mL (about 450mg).After fat grafting 3 months, outside graft tunicle, separate graft, investigate and move
Vegetable fat fat tissue survival rate and fibrosis and degree of necrosis.
(1) graft weight in wet base
Graft is carried out weight in wet base mensuration.Result is as shown in table 4.
Table 4, the weight in wet base statistics of graft
| Group | Weight in wet base |
| Matched group | 92.76±6.70mg |
| Experimental group 1 | 134.58±7.67mg* |
| Experimental group 2 | 146.46±8.46mg* |
| Experimental group 3 | 164.89±7.58mg* |
| Experimental group 4 | 186.73±11.42mg* |
| Experimental group 5 | 205.57±12.67mg* |
* represent that compared with matched group P < 0.05 has significant difference.
Statistic software SPSS 17.0 is used to carry out data process the data of table 4.Use the side of randomized block design data
Difference analysis, compares weight in wet base.Knowable to the data of table 4, the graft weight in wet base that matched group obtains is about 92.67mg, real
The graft weight in wet base testing group 1 is about 134.58mg, and the graft weight in wet base of experimental group 2 is about 146.46mg, the graft of experimental group 3
Weight in wet base about 164.89mg, the graft weight in wet base of experimental group 4 are 187.73mg, the graft weight in wet base of experimental group 5 is 205.57mg,
Each experimental group, compared with matched group, is respectively provided with significant difference, P < 0.05.
Additionally, knowable to the data of experimental group 1～experimental group 5, along with umbilical cord mesenchymal stem cells addition and cell because of
The increase of sub-PDGF, graft weight in wet base also increases, and presents positively related relation.
(2) fibrosis of transplant fat and degree of necrosis
Use " some counting " measuring method that Gundersen proposed in 1998.Use grid software test system, including 20
Line and 100 intersection points, measure 10 200 times of visuals field (two with blind extraction not repeating mutually, randomly choosing of every section
Observer measures, the fibrosis in l0 the visual field of counting and the number of blister cavities), the some number that each group of graft measurement is obtained
Carry out the comparison of fibrosis and degree of necrosis.Result is as shown in table 5.
Table 5, fibrosis and degree of necrosis result table
| Group | Point system (fibrosis and necrosis) |
| Matched group | (650.5 soil 4.5) individual/IOHF |
| Experimental group 1 | (316.5 soil 4.5) individual/IOHF* |
| Experimental group 2 | (220.2 soil 5.8) individual/IOHF* |
| Experimental group 3 | (215.5 soil 8.5) individual/IOHF* |
| Experimental group 4 | (150.5 soil 3.5) individual/IOHF* |
| Experimental group 5 | (108.5 soil 7.5) individual/IOHF* |
Statistic software SPSS 17.0 is used to carry out data process.Use the variance analysis of randomized block design data, to fibre
Dimensionization and blister cavities number row compare.
Result shows, without any component in matched group, fibrosis and the degree of necrosis of its graft are the highest, is about
650.5 individual/IOHF；Experimental group 1 is about 316.5/IOHF, and experimental group 2 is about 220.2/IOHF, experimental group 3 is about 215.5
Individual/IOHF, experimental group 4 are about 150.5/IOHF, experimental group 5 is about 108.5/IOHF, each experimental group compared with matched group,
Experimental group 1～5 groups of transplant fat tissue fibering and blister cavities number are the most less, are respectively provided with significant difference, P < 0.05.
Embodiment described above only have expressed the several embodiments of the present invention, and it describes more concrete and detailed, but also
Therefore the restriction to the scope of the claims of the present invention can not be interpreted as.It should be pointed out that, for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, it is also possible to make some deformation and improvement, these broadly fall into the guarantor of the present invention
Protect scope.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. a fatty tissue compound formulation, it is characterised in that: comprise platelet derived growth factor, umbilical cord mesenchymal stem cells
And stromal vascular component.
Fatty tissue compound formulation the most according to claim 1, it is characterised in that: possibly together with fatty tissue.
Fatty tissue compound formulation the most according to claim 1, it is characterised in that: blood in described fatty tissue compound formulation
The content of platelet derivative growth factor is 5～20ng/ml.
Fatty tissue compound formulation the most according to claim 1, it is characterised in that: umbilicus in described fatty tissue compound formulation
Quantity with mescenchymal stem cell is 5 × 105～5 × 106Individual/ml.
Fatty tissue compound formulation the most according to claim 1, it is characterised in that: base in described fatty tissue compound formulation
The content of matter vascular component is 5 × 105～5 × 106Individual/ml.
Fatty tissue compound formulation the most according to claim 2, it is characterised in that: described fatty tissue compound formulation contains
The following component of following content:
Umbilical cord mesenchymal stem cells 1 × 106～5 × 106Individual/ml,
Stromal vascular component 1 × 106Individual/ml,
Platelet derived growth factor 5～20ng/ml.
Fatty tissue compound formulation the most according to claim 6, it is characterised in that: described fatty tissue compound formulation contains
The following component of following content:
Umbilical cord mesenchymal stem cells 1 × 106Individual/ml,
Stromal vascular component 1 × 106Individual/ml,
Platelet derived growth factor 10ng/ml.
8. the preparation method of fatty tissue compound formulation described in claim 6 or 7, comprises the following steps: by stromal vascular component,
Umbilical cord mesenchymal stem cells, platelet derived growth factor are configured to cell suspension, then by fatty tissue and cell suspension according to
The ratio mixing of volume ratio 20:1.
9. fatty tissue compound formulation described in any one of claim 2-7 shaping or beauty treatment in as filling preparation application.
Application the most according to claim 9, it is characterised in that: described shaping or beauty treatment include the depression of filling facial area
Deformity, mamaplasty, rich temporo and augmentation rhinoplasty.
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| WO2019161540A1 (en) * | 2018-02-23 | 2019-08-29 | 高扬 | Pluripotent stem cell differentiation and regeneration-based cosmetic preparation for delaying aging of body |
| 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 |
| WO2021237900A1 (en) * | 2020-05-28 | 2021-12-02 | 澳门大学 | Application of mesenchymal stem cell in preparing preparation for promoting fat grafting |
| CN115605235A (en) * | 2020-03-22 | 2023-01-13 | 科弗朗有限公司（Il） | Collagen-based formulations useful as soft tissue fillers and/or implants |
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