CN106727705A - A kind of stem cell medicine and its preparation method and application - Google Patents
A kind of stem cell medicine and its preparation method and application Download PDFInfo
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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Abstract
The present invention relates to stem cell medicine technical field, more particularly to a kind of stem cell medicine and its preparation method and application.The stem cell medicine includes fat mesenchymal stem cell, platelet rich plasma and hyaluronic acid aqueous solution.After be applied in combination for fat mesenchymal stem cell and PRP, hyaluronic acid or its salt by the present invention, there is good repair to articular cartilage damage, effect is significant is better than micro fractures operation.
Description
Technical field
The present invention relates to stem cell medicine technical field, more particularly to a kind of stem cell medicine and preparation method thereof and should
With.
Background technology
Articular cartilage is a kind of viscoelastic materials, and different moving parts show different load-bearer properties.Articular cartilage
This key property and its reduce articular surface friction effect be cartilage ultra microstructure composition and chemical component basic training
Can embody, articular cartilage is formed by sparse cell arrangement, without blood vessel, lymph and nerve supply.Articular cartilage is in joint motion
In play an important role, its structure is very fine, science, to adapt to different function needs.Wound or injury gained in sports often result in pass
The acute injury of cartilage is saved, causes arthralgia, swelling and dysfunction, have a strong impact on the quality of life of patient.Cartilage damage
Do not have afterwards or few self-repairing capabilities.Therefore repair of cartilage turns into field of orthopaedics major issue.
Micro fractures are performed the operation, i.e., select several points in diseased region, are drilled to subchondral bone, make defect and subchondral bone
Marrow is communicated, and the mesenchymal stem cells MSCs (mesenchymal stem cells, MSCs) with differentiation potential can be oozed out
Into defective region, the very strong stem cell cartilage surface for being covered in defect mixed in together of blood and power of regeneration in marrow is carried out
Repair of cartilage.Cartilage premature wear can carry out regenerating bone or cartilage art.Full-thickness cartilage and subchondral bone are damaged certain reparation energy
Power, part cartilage damage is drilled to subchondral bone is inducible and is repaired, and repair tissue has osteogenic ability in originating from cancellous bone
Mescenchymal stem cell, repair tissue be class hyaline cartilage.But the method for micro fractures operation need to be performed the operation and be beaten in diseased region
Hole, patient bears very big pain, and with certain risk, and MSCs amounts in marrow are little, generate the fiber of prosthetic
Time needed for cartilaginous tissue is more long.
With the development of biotechnology, research of the stem cell in various disease models are treated is more and more more, because it has
There is self-renewing and polytype cell can be divided into, have a wide range of applications, therefore also by soft as treatment joint
The main study subject of bone injury.But directly by stem cell injection treatment articular cartilage damage exist cell survival rate it is relatively low,
The difficult shortcoming of cell propagation.Therefore, it is badly in need of providing a kind of pharmaceutical preparation that can effectively treat articular cartilage damage.
The content of the invention
In view of this, the invention provides a kind of stem cell medicine and its preparation method and application.In the stem cell medicine
Stem cells hyperplasia activity is good, can effectively treat articular cartilage damage.
In order to realize foregoing invention purpose, the present invention provides following technical scheme：
The invention provides a kind of stem cell medicine, including fat mesenchymal stem cell, platelet rich plasma and hyalomitome
Aqueous acid.
Fat mesenchymal stem cell (adipose-derived stem cells, ADSCs) is in recent years from adipose tissue
In a kind of isolated mescenchymal stem cell with multi-lineage potential.Research finds that ADSCs cells can be steady in vitro
Breed calmly and decline rate is low, while there are materials to organize easily, on a small quantity for it can obtain a large amount of stem cells, it is suitable to train on a large scale
Support, the advantages of small to body injury, and its wide material sources, greatly, suitable autotransplantation is increasingly becoming in recent years cylinder storage amount
One of new study hotspot.
Platelet rich plasma (PRP) refer to using the blood of itself, extract rich in high concentration blood platelet and it is various from
The blood plasma of body growth factor.PRP plays positive effect in rush wound healing, activation tissue in situ Stem Cell Activity, pre- through PRP
The MSCs for the treatment of is transplanted to survival rate after body and improves, and the biological agent enhancing such as its angiogenesis promoting.
Hyaluronic acid is the main component of knuckle synovia, can prevent or delay further regression, clearly with Saving cortilage cartilage
Except algogenic substance, play the role of substantially to mitigate pain, can be used for treatment of arthritis and accelerating wound healing.
After be applied in combination for fat mesenchymal stem cell and PRP, hyaluronic acid or its salt by the present invention, to articular cartilage damage
With good repair, effect is significant is better than micro fractures operation, and fat mesenchymal stem cell convenient material drawing, proliferation activity
Well, purity is high, can in a short time obtain a fairly large number of MSCs, and the stem cell medicine can mitigate the treatment pain and wind of patient
Danger, strengthens curative effect, shortens treatment time.
Preferably, the consumption of each component is in the stem cell medicine：
Fat mesenchymal stem cell：(1~10) × 106It is individual；
Platelet rich plasma：1~10mL；
Hyaluronic acid aqueous solution：1~10mL.
In the embodiment that the present invention is provided, the consumption of each component is in stem cell medicine：
Fat mesenchymal stem cell：(1~3) × 106It is individual；
Platelet rich plasma： 2mL；
Hyaluronic acid aqueous solution： 2mL.
Preferably, the mass percentage concentration of hyaluronic acid aqueous solution is 0.0008~0.0012g/mL.
In the present invention, hyaluronic acid can also be hyaluronate, such as Sodium Hyaluronate, can equally realize this hair
It is bright.In the embodiment that the present invention is provided, hyaluronate is Sodium Hyaluronate.
In the embodiment that the present invention is provided, fat mesenchymal stem cell is 2nd generation fat mesenchymal stem cell.
In the embodiment that the present invention is provided, the formulation of stem cell medicine is injection.
Present invention also offers application of the stem cell medicine in repair of cartilage medicine is prepared.
In the embodiment that the present invention is provided, cartilage is articular cartilage.
Present invention also offers the preparation method of the stem cell medicine, including：It is dry thin using the resuspended fat mesenchymals of PRP
Born of the same parents, then mix with hyaluronic acid aqueous solution, obtain the stem cell medicine.
In the embodiment that the present invention is provided, the preparation method of fat mesenchymal stem cell is：By adipose tissue physiology
Normal saline washing, removes the haemocyte and fragment of tissue of residual.To 0.1% isometric NTx enzyme is added in tissue, 37 are put into
30min is digested in DEG C gas bath oscillator.Then with 1500r/min, upper-layer fat tissue and lower floor's suspension are discarded after centrifugation 10min
Obtain ADSCs.Its is resuspended, and density is adjusted to 1 × 105/ mL, is then seeded in culture area for 25cm2Tissue Culture Flask
In.When ADSCs degrees of fusion up to 80% or so, collect cell using, pass on or freeze.
In the embodiment that the present invention is provided, the preparation method of platelet rich plasma is：Venous puncture whole blood；For the first time from
The heart, 1600r/min is centrifuged 10min, and blood will abandon red blood cell layer by 2 layers of layering blood plasma and red blood cell；It is centrifuged for second,
5000r/min, is centrifuged 5min, is inhaled after centrifugation and abandons part blood plasma, with the blood of 1/2nd volumes of remaining preparation final volume
Resuspended precipitation is starched, PRP is prepared into.
The invention provides a kind of stem cell medicine and its preparation method and application.The stem cell medicine fills between including fat
Matter stem cell, platelet rich plasma and hyaluronic acid aqueous solution.The present invention at least has one of following advantage：
1st, after be applied in combination for fat mesenchymal stem cell and PRP, hyaluronic acid or its salt by the present invention, articular cartilage is damaged
Wound has good repair, and effect is significant is better than micro fractures operation；
2nd, fat mesenchymal stem cell convenient material drawing：Liposuction is simple to operate, and pain is few, need to only extract a small amount of fat group
Knitting can just turn out substantial amounts of fat stem cell, and only extract once fat, it is possible to preparation of preparation several times；Fat mesenchymal
Stem cells hyperplasia activity is good, purity is high, can in a short time obtain a fairly large number of MSCs, generates the fibrocartilage group of prosthetic
The better efficacy knitted；
3rd, stem cell medicine of the present invention can mitigate the treatment pain and risk of patient using simple to operate, strengthen curative effect, contracting
Short treatment time.
Brief description of the drawings
Fig. 1 shows the form of P2 fat subsitutes mescenchymal stem cells, wherein, 1-1 shows that P2 amplifies 40 times of form, 1-2 for cell
Show that P2 amplifies 100 times of form for cell.
Specific embodiment
The invention discloses a kind of stem cell medicine and its preparation method and application, those skilled in the art can use for reference this
Literary content, is suitably modified technological parameter realization.In particular, all similar replacements and change are to art technology
It is it will be apparent that they are considered as being included in the present invention for personnel.The method of the present invention and application have passed through preferably
Embodiment is described, and related personnel can be not substantially being departed from present invention, spirit and scope to side as herein described
Method and application are modified or suitably change is realized and apply the technology of the present invention with combining.
The invention provides a kind of stem cell medicine, its preparation method includes：
1) ADSCs cultures
The adipose tissue normal saline flushing that liposuction is originated, removes the haemocyte and fragment of tissue of residual.To tissue
It is middle to add isometric 0.1% NTx enzyme, it is put into 37 DEG C of gas bath oscillators and digests 30min.Then with 1500r/min, from
Upper-layer fat tissue is discarded after heart 10min and lower floor's suspension obtains ADSCs.Its is resuspended, and density is adjusted to 1 × 105/ mL, so
After be seeded in culture area for 25cm2Tissue Culture Flask in.When ADSCs degrees of fusion up to 80% or so, collect cell and use, pass
In generation, freezes.
2) PRP is prepared
After cleaning-sterilizing, 10~20ml of venous puncture whole blood.It is centrifuged for the first time, 1600r/min, 10min is centrifuged, blood will
By 2 layers of layering blood plasma and red blood cell, red blood cell layer is abandoned；It is centrifuged for second, 5000r/min is centrifuged 5min, is inhaled after centrifugation and abandon portion
Divide blood plasma, with the resuspended precipitation of blood plasma of 1/2nd volumes of remaining preparation final volume, be prepared into PRP.
3) cultured ADSCs (1~3) × 10 in taking 1)6It is individual, with the resuspended stem cells of 2ml PRP for 2) preparing.
4) sodium hyaluronate solution is taken, is mixed in equal volume with the PRP containing stem cell, be prepared into final finished preparation, preparation
Middle sodium hyaluronate solution is 1 with PRP volume ratios:1, stem cell does not account for volume.
5) to intraarticular injection preparation.
Experimental animal used, reagent or instrument in stem cell medicine that the present invention is provided and its preparation method and application
Bought by market.In the embodiment that the present invention is provided, the mass percentage concentration of sodium hyaluronate solution is 0.001g/mL.It is complete
Full culture medium is the DMEM-F12 culture mediums containing 10%FBS.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1
(1) ADSCs cultures
2-3 weeks in advance, the rabbit adipose tissue about 10ml normal saline flushings that liposuction is originated removed the haemocyte of residual
And fragment of tissue.To 0.1% isometric NTx enzyme is added in tissue, it is put into 37 DEG C of gas bath oscillators and digests 30min.
Then with 1500r/min, discard upper-layer fat tissue after centrifugation 10min and lower floor's suspension obtains ADSCs.Its is resuspended, density
It is adjusted to 1 × 105/ mL, is then seeded in culture area for 25cm2Tissue Culture Flask in.Treat that cell growth to degree of converging reaches
80%, after washing 2 times with PBS, 0.015ml/cm is added toward cell20.25% pancreatin+0.04%EDTA digestion 2min, use
10 times of complete mediums of digestive juice terminate enzymolysis, and sampling is counted, 200g centrifugation 5min, remove supernatant, according to count results, use
It is 5 × 10 that complete medium re-suspended cell is precipitated to cell density4Cell/mL, inoculated and cultured.When ADSCs degrees of fusion up to 80%
Left and right, collect cell using, pass on or freeze.P2 is shown in Fig. 1 for ADSCs cellular morphology pictures.
As seen from Figure 1, passage cell is observed under inverted microscope, multiplication capacity is stronger, the speed of growth is very fast, it is form, big
It is small homogeneous, it is radial to the growth of fusiformis proper alignment.Before freezing, cell refractivity is strong, in spindle shape, without heteroproteose cell, background
Clearly, cell purity is higher.
(2) PRP is prepared
After cleaning-sterilizing, venous puncture whole blood 20ml.It is centrifuged for the first time, 1600r/min, 10min is centrifuged, blood will be divided
2 layers of layer blood plasma and red blood cell, abandon red blood cell layer；It is centrifuged for second, 5000r/min is centrifuged 5min, is inhaled after centrifugation and abandon part-blood
Slurry, with the resuspended precipitation of remaining 2ml or so blood plasma, is prepared into PRP.
(3) preparation of stem cell medicine
Cultured P2 is for ADSCs cells 1 × 10 in taking (1)6It is individual, with the resuspended ADSCs of 2mL PRP.
2mL sodium hyaluronate solutions are taken, is mixed in equal volume with the PRP containing ADSCs, be prepared into final finished preparation.
(4) effect identification
1) rabbit cartilage damage model is set up
The back leg knee joint inner side stringer holostrome of rabbit is cut under gnotobasis, total length 2cm, go directly articular cavity, hand drill
A diameter thickness up to the circular opening of cartilage holostrome is bored on front side of femoral intercondylar, holostrome knee articular cartilage defect animal mould is caused
Type, otch interrupted whole sutures four or five pins.The postoperative equal intramuscular injection gentamicin 4 × 10 of each experimental animal4U/d to postoperative 3rd day with
The POI that prevention is likely to occur.Experimental animal is put back to cage and normally raises, not brake measure.Take 9 rabbits at random
It is divided into 3 groups, every group 3.
2) experiment packet
1 group of comparative example：Using arthroscope, in the subchondral plate punching manufacture micro fractures of cartilaginous lesion site, when seeing bone
When marrow blood comes out from hole, illustrate that punching depth has reached requirement.Take 2ml sodium hyaluronate solutions abundant with 2ml PRP
After mixing, it is expelled in the articular cavity of the rabbit for having carried out micro fractures operation.(micro fractures operation+PRP+ Sodium Hyaluronates)
2 groups of comparative example：Take after 2ml sodium hyaluronate solutions and 2ml PRP fully mix, be expelled to the articular cavity of rabbit
It is interior.(PRP+ Sodium Hyaluronates)
3 groups of comparative example：P2 is taken for ADSCs cells 1 × 106It is individual, with the resuspended ADSCs of 4mL PRP.
4 groups of comparative example：P2 is taken for ADSCs cells 1 × 106It is individual, with the resuspended ADSCs of 4mL sodium hyaluronate solutions.
1 group of embodiment：The final finished preparation that the present embodiment 1 is prepared into is expelled in the articular cavity of rabbit.(ADSCs+
PRP+ Sodium Hyaluronates)
3) result of the test
Animal general status are observed：9 rabbits it is postoperative it is equal normal activity, feed and can drink water, do not occur knee joint it is red and swollen,
The infection performance such as suppurate, other diseases situation do not occur.
Repair of cartilage general form observes result：3 months after operation, puts to death each group experimental rabbit respectively, and knee cartilage is entered
Row general form is observed.It is specific as follows：
The damaged cartilage region of embodiment 1 is not obvious, all by chondroid tissue covering, cambium and surrounding normal cartilage
Tissue is without obvious boundary, and surface is smooth；
The articular cartilage damage region of comparative example 1 is more apparent, edge hyperplasia, form rule, central portion out-of-flatness, it is seen that newborn
Tissue coverage, covering tissue is white, unclear with damage margo, and cartilage degeneration is more serious.
Substantially, edge substantially, has no or seldom sees the damaged cartilage region of comparative example 2 without hyperplasia, form irregular, crack
Cambium is covered, and cartilage degeneration is serious.
The articular cartilage damage region of comparative example 3 is less obvious, it is seen that cambium is covered, and covering tissue is white, with damage
Hinder margo to obscure.
The articular cartilage damage region of comparative example 4 is less obvious, it is seen that cambium is covered, and covering tissue is white, with damage
Hinder margo to obscure.
Embodiment 2
(1) ADSCs cultures
2-3 weeks in advance, the rabbit adipose tissue about 10ml normal saline flushings that liposuction is originated removed the haemocyte of residual
And fragment of tissue.To 0.1% isometric NTx enzyme is added in tissue, it is put into 37 DEG C of gas bath oscillators and digests 30min.
Then with 1500r/min, discard upper-layer fat tissue after centrifugation 10min and lower floor's suspension obtains ADSCs.Its is resuspended, density
It is adjusted to 1 × 105/ mL, is then seeded in culture area for 25cm2Tissue Culture Flask in.Treat that cell growth to degree of converging reaches
80%, after washing 2 times with PBS, 0.015ml/cm is added toward cell20.25% pancreatin+0.04%EDTA digestion 2min, use
10 times of complete mediums of digestive juice terminate enzymolysis, and sampling is counted, 200g centrifugation 5min, remove supernatant, according to count results, use
It is 5 × 10 that complete medium re-suspended cell is precipitated to cell density4Cell/mL, inoculated and cultured.When ADSCs degrees of fusion up to 80%
Left and right, collect cell using, pass on or freeze.
(2) PRP is prepared
After cleaning-sterilizing, venous puncture whole blood 20ml.It is centrifuged for the first time, 1600r/min, 10min is centrifuged, blood will be divided
2 layers of layer blood plasma and red blood cell, abandon red blood cell layer；It is centrifuged for second, 5000r/min is centrifuged 5min, is inhaled after centrifugation and abandon part-blood
Slurry, with the resuspended precipitation of remaining 2ml or so blood plasma, is prepared into PRP.
(3) preparation of stem cell medicine
Cultured P2 is for ADSCs cells 2 × 10 in taking (1)6It is individual, with the resuspended ADSCs of 2mL PRP.
2mL sodium hyaluronate solutions are taken, is mixed in equal volume with the PRP containing ADSCs, be prepared into final finished preparation.
(4) effect identification
1) rabbit cartilage damage model is set up
The right side rear knee joint inner side stringer holostrome of rabbit is cut under gnotobasis, total length 2cm, go directly articular cavity, hand
Brace bores a diameter thickness up to the circular opening of cartilage holostrome on front side of femoral intercondylar, causes holostrome knee articular cartilage defect animal
Model, otch interrupted whole sutures four or five pins.The postoperative equal intramuscular injection gentamicin 4 × 10 of each experimental animal4U/d to postoperative 3rd day
With the POI for preventing to be likely to occur.Experimental animal is put back to cage and normally raises, not brake measure.Take 3 rabbits at random
Son.
2) result of the test
Animal general status are observed：, there is not knee joint redness, stream in the postoperative equal energy normal activity of rabbit, feed and drinking-water
, there is not other diseases situation in the infection performance such as purulence.
Repair of cartilage general form observes result：3 months after operation, puts to death experimental rabbit, and the big bodily form is carried out to knee cartilage
State is observed.The damaged cartilage region of embodiment 2 is not obvious, all by chondroid tissue covering, cambium and surrounding normal cartilage
, without obvious boundary, surface is smooth, does not have notable difference with the Contrast on effect of embodiment 1 for tissue.
Embodiment 3
(1) ADSCs cultures
2-3 weeks in advance, the rabbit adipose tissue about 10ml normal saline flushings that liposuction is originated removed the haemocyte of residual
And fragment of tissue.To 0.1% isometric NTx enzyme is added in tissue, it is put into 37 DEG C of gas bath oscillators and digests 30min.
Then with 1500r/min, discard upper-layer fat tissue after centrifugation 10min and lower floor's suspension obtains ADSCs.Its is resuspended, density
It is adjusted to 1 × 105/ mL, is then seeded in culture area for 25cm2Tissue Culture Flask in.Treat that cell growth to degree of converging reaches
80%, after washing 2 times with PBS, 0.015ml/cm is added toward cell20.25% pancreatin+0.04%EDTA digestion 2min, use
10 times of complete mediums of digestive juice terminate enzymolysis, and sampling is counted, 200g centrifugation 5min, remove supernatant, according to count results, use
It is 5 × 10 that complete medium re-suspended cell is precipitated to cell density4Cell/mL, inoculated and cultured.When ADSCs degrees of fusion up to 80%
Left and right, collect cell using, pass on or freeze.
(2) PRP is prepared
After cleaning-sterilizing, venous puncture whole blood 20ml.It is centrifuged for the first time, 1600r/min, 10min is centrifuged, blood will be divided
2 layers of layer blood plasma and red blood cell, abandon red blood cell layer；It is centrifuged for second, 5000r/min is centrifuged 5min, is inhaled after centrifugation and abandon part-blood
Slurry, with the resuspended precipitation of remaining 2ml or so blood plasma, is prepared into PRP.
(3) preparation of stem cell medicine
Cultured P2 is for ADSCs cells 3 × 10 in taking (1)6It is individual, with the resuspended ADSCs of 2mL PRP.
2mL sodium hyaluronate solutions are taken, is mixed in equal volume with the PRP containing ADSCs, be prepared into final finished preparation.
(4) effect identification
1) rabbit cartilage damage model is set up
The right side rear knee joint inner side stringer holostrome of rabbit is cut under gnotobasis, total length 2cm, go directly articular cavity, hand
Brace bores a diameter thickness up to the circular opening of cartilage holostrome on front side of femoral intercondylar, causes holostrome knee articular cartilage defect animal
Model, otch interrupted whole sutures four or five pins.The postoperative equal intramuscular injection gentamicin 4 × 10 of each experimental animal4U/d to postoperative 3rd day
With the POI for preventing to be likely to occur.Experimental animal is put back to cage and normally raises, not brake measure.Take 3 rabbits at random
Son.
2) result of the test
Animal general status are observed：, there is not knee joint redness, stream in the postoperative equal energy normal activity of rabbit, feed and drinking-water
, there is not other diseases situation in the infection performance such as purulence.
Repair of cartilage general form observes result：3 months after operation, puts to death experimental rabbit, and the big bodily form is carried out to knee cartilage
State is observed.The damaged cartilage region of embodiment 3 is not obvious, all by chondroid tissue covering, cambium and surrounding normal cartilage
, without obvious boundary, surface is smooth, does not have notable difference with the Contrast on effect of embodiment 1 for tissue.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of stem cell medicine, it is characterised in that including fat mesenchymal stem cell, platelet rich plasma and hyalomitome sour water
Solution.
2. stem cell medicine according to claim 1, it is characterised in that the consumption of each component in the stem cell medicine
For：
Fat mesenchymal stem cell：(1~10) × 106It is individual；
Platelet rich plasma：1~10mL；
Hyaluronic acid aqueous solution：1~10mL.
3. stem cell medicine according to claim 1 and 2, it is characterised in that the use of each component in the stem cell medicine
Measure and be：
Fat mesenchymal stem cell：(1~3) × 106It is individual；
Platelet rich plasma： 2mL；
Hyaluronic acid aqueous solution： 2mL.
4. stem cell medicine according to any one of claim 1 to 3, it is characterised in that the hyaluronic acid aqueous solution
Mass percentage concentration be 0.0008~0.0012g/mL.
5. stem cell medicine according to any one of claim 1 to 4, it is characterised in that the hyaluronate is
Bright matter acid sodium.
6. stem cell medicine according to any one of claim 1 to 5, it is characterised in that the fat mesenchymal is dry thin
Born of the same parents are 2nd generation fat mesenchymal stem cell.
7. stem cell medicine according to any one of claim 1 to 6, it is characterised in that the agent of the stem cell medicine
Type is injection.
8. application of the stem cell medicine in repair of cartilage medicine is prepared as any one of claim 1 to 7.
9. application according to claim 8, it is characterised in that the cartilage is articular cartilage.
10. as any one of claim 1 to 7 stem cell medicine preparation method, it is characterised in that including：Using richness
The resuspended fat mesenchymal stem cell of thrombocyte plasma, then mixes with hyaluronic acid aqueous solution, obtains the stem cell medicine.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107349468A (en) * | 2017-07-02 | 2017-11-17 | 江西瑞济生物工程技术股份有限公司 | A kind of amnion stem cell gel and its preparation method and application |
| CN110226591A (en) * | 2019-06-14 | 2019-09-13 | 西安飞如生物科技有限公司 | Cell-preservation liquid and its application and fibroblast injection |
| CN110846274A (en) * | 2019-11-20 | 2020-02-28 | 广东先康达生物科技有限公司 | Preparation method of compound cell preparation applied to osteoarthritis and use method of compound cell preparation |
| CN111454901A (en) * | 2020-05-09 | 2020-07-28 | 山东兴瑞生物科技有限公司 | Method for inducing differentiation of adipose-derived stem cells into chondrocytes |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103071191A (en) * | 2013-02-04 | 2013-05-01 | 成都清科生物科技有限公司 | Preparation method of autologous platelet-factor-rich plasma (PFRP) preparation |
| CN104958320A (en) * | 2015-07-29 | 2015-10-07 | 西安芙金细胞科技有限公司 | Cell preparation for treating osteoarthritis and preparation method thereof |
| CN105168251A (en) * | 2015-09-09 | 2015-12-23 | 广州赛莱拉干细胞科技股份有限公司 | Stem cell preparation as well as preparation method and application thereof |
-
2016
- 2016-12-26 CN CN201611217293.4A patent/CN106727705A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103071191A (en) * | 2013-02-04 | 2013-05-01 | 成都清科生物科技有限公司 | Preparation method of autologous platelet-factor-rich plasma (PFRP) preparation |
| CN104958320A (en) * | 2015-07-29 | 2015-10-07 | 西安芙金细胞科技有限公司 | Cell preparation for treating osteoarthritis and preparation method thereof |
| CN105168251A (en) * | 2015-09-09 | 2015-12-23 | 广州赛莱拉干细胞科技股份有限公司 | Stem cell preparation as well as preparation method and application thereof |
Cited By (6)
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|---|---|---|---|---|
| CN107349468A (en) * | 2017-07-02 | 2017-11-17 | 江西瑞济生物工程技术股份有限公司 | A kind of amnion stem cell gel and its preparation method and application |
| CN110226591A (en) * | 2019-06-14 | 2019-09-13 | 西安飞如生物科技有限公司 | Cell-preservation liquid and its application and fibroblast injection |
| CN110226591B (en) * | 2019-06-14 | 2021-05-18 | 西安飞如生物科技有限公司 | Cell preservation solution, application thereof and fibroblast injection |
| CN110846274A (en) * | 2019-11-20 | 2020-02-28 | 广东先康达生物科技有限公司 | Preparation method of compound cell preparation applied to osteoarthritis and use method of compound cell preparation |
| CN111454901A (en) * | 2020-05-09 | 2020-07-28 | 山东兴瑞生物科技有限公司 | Method for inducing differentiation of adipose-derived stem cells into chondrocytes |
| CN111454901B (en) * | 2020-05-09 | 2022-07-12 | 山东兴瑞生物科技有限公司 | Method for inducing differentiation of adipose-derived stem cells into chondrocytes |
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