CN104383726A - Extraction method of platelet rich plasma and extracted platelet rich plasma - Google Patents
Extraction method of platelet rich plasma and extracted platelet rich plasma Download PDFInfo
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Abstract
The invention relates to an extraction method of a platelet rich plasma and an extracted platelet rich plasma. Particularly, the method for extracting the platelet rich plasma from blood comprises the following steps: (a) putting collected whole blood in a container with an anticoagulant, and sufficiently mixing the blood and the anticoagulant; (b) putting the blood mixed with the anticoagulant in a centrifugal pipe for first-time centrifugation, and dividing the blood into three layers basically; (c) drawing the topmost layer and most of the middle layer of blood and transferring the blood to a new centrifugal pipe, and uniformly mixing for second-time centrifugation; (d) removing the plasma on the upper layer of the centrifugal pipe, and suspending deposited blood platelets again by using the rest of plasma to obtain the platelet rich plasma (PRP). The method is simple in operation process and high in yield, and the concentration of the blood platelets in the enriched plasma is high.
Description
The application is the Chinese Patent Application No. 201210267055X submitted on July 30th, 2012, and denomination of invention is the divisional application of " extracting method of platelet rich plasma and the platelet rich plasma of extraction " case, and its full content is incorporated herein by reference.
Technical field
The invention belongs to technical field of clinical medicine, be specifically related to a kind of method that hematoblastic blood plasma is rich in extraction, the platelet rich plasma that the inventive method obtains under given conditions has that PC is high, the advantage of recovery rate of blood platelet at least one aspect high.
Background technology
Be rich in hematoblastic blood plasma, can be described as platelet rich plasma (Platelet Rich Plasma, PRP) in the present invention, it is the thrombocyte plasma containing high concentration obtained by centrifugal blood.The blood platelet of human body is except except providing cohesion when hemostasis, can also contain a lot of multiple growth factor relevant with wound healing and bone regeneration in blood platelet.As platelet derived growth factor (Platelet Derived Growth Factor, PDGF), TGF (Transforming Growth Factor, TGF), IGF (Insulin-like Growth Factor, IGF), EGF (Epidermal Growth Factor, and VEGF (Vascular Endothelial Growth Factor EGF), VEGF) etc., the growth factor in blood platelet source, facilitation is played to the Differentiation and proliferation of cell, these growth factors have cooperative response each other, factor interaction and the impact of cytoactive is promoted with other, jointly maintain the balance of organizational environment, to wound healing, repair and regenerate and have important effect.
In addition, because PRP can extract from patient's self-blood, after Extraction and enrichment process, can be used for the treatment of relevant disease, thus abundance, draw materials conveniently, preparation method is simple and can allow body absorption.Self PRP uses and the problem of virus propagation and immunological rejection can be avoided to occur.Therefore PRP is widely applied in different medical domains.As orthopaedics, the department of stomatology, decorative sursery, sports medical science and aesthetic medicine.
PRP is going back the unified preparation method of neither one so far, and the preparation principle of PRP mainly utilizes various composition sinking speed in blood different, by centrifugal by blood layering.Thus obtain containing hematoblastic blood plasma, and utilize centrifugal principle further concentrated with the thrombocyte plasma obtaining high concentration.In human whole blood, hematoblastic concentration is generally 1 ~ 3 × 10
5/ ml.Research thinks that the PRP PC after concentrating should be the 3-4 of Whole blood platelet concentration doubly.Research also finds that the raising of PC in PRP effectively can improve the propagation of stem cell and differentiation capability and significantly can increase the expression of fibroblastic propagation and I-type collagen.
Therefore be effectively separated and improve hematoblastic concentration in PRP and become the key of whole PRP therapeutic scheme.
(the CN101402940A such as Lin Shuru, Chinese Patent Application No.: 200810228726.5, denomination of invention: a kind of method for extracting plastocyte of mouse) disclose a kind of method for extracting plastocyte of mouse, mouse socket of the eye venous blood sampling, be placed in anti-freezing centrifuge tube, centrifugal after leaving standstill 30MIN, the centrifugal 10MIN of 800RPM, supernatant is and is rich in hematoblastic blood plasma.Sucking-off supernatant is placed in clean tube, the centrifugal 10MIN of 3500RPM, abandoning supernatant, and sediment at the bottom of pipe is blood platelet.In test tube, drip 50 ~ 100 Μ L mass concentrations is 1% ammonium oxalate solution, stirs gently with glass bar, then to add mass concentration be 1% ammonium oxalate solution 2 ~ 3ML, leaves standstill 5MIN, makes erythrocytolysis.The centrifugal 10MIN of 3500RPM, supernatant discarded, adds blood platelet cleaning solution and repeatedly blows and beats, making to become thrombocyte suspension, the centrifugal 10MIN of 3300RPM, supernatant discarded, adds a small amount of blood platelet cleaning solution and repeatedly blow and beat to make it to become suspension.This blood platelet as after antigen immune cavy, will get GPS to injected in mice, preparation anaphylactoid purpura model.The method is simple to operate, can obtain the blood platelet of high concentration, and solves the not good problem of blood platelet purity well.Although this method of carrying out platelet rich for mouse blood it is believed that can obtain concentration reaches 10
10blood platelet, but the method its hematoblastic yield unexposed, and method is operationally more numerous and diverse, is difficult to the process being applicable to human blood.
In addition, (CN 102078644 A such as Wang Yue, Chinese Patent Application No. 201110053979.5, denomination of invention: a kind of autologous platelet rich plasma extraction element of simple and effective and extracting method) in disclose a kind of autologous platelet rich plasma extraction element and extracting method of simple and effective, containing multiple composite growth factor in the autologous platelet rich plasma extracted, the reparation of tissue effectively can be promoted.Be made up of 1 syringe, 1 intravenous transfusion device tube connector, 1 venous detaining needle plastic cards and 1 intravenous infusion apparatus needle; Syringe is in order to venous blood samples and serve as centrifuge tube; Intravenous infusion apparatus needle is connected with syringe outlet by intravenous transfusion device tube connector, intravenous infusion apparatus needle in order to skin heart puncture, extract anti-coagulants and activator and need the autologous platelet rich plasma after injection location extraction to human body; Intravenous transfusion device tube connector is flexible to be fixed in venous detaining needle plastic cards.It is believed that this device is useful in the operability promoting blood platelet extraction.
Therefore in order to enrichment blood platelet and obtain the platelet rich plasma of clinical treatment meaning from blood, seeking that a kind of operating process is simple, yield is high, PC is high in the blood plasma of enrichment method, is that those skilled in the art urgently expect.
Summary of the invention
The object of the invention is to explore a kind of simple and hematoblastic method in isolation and identification blood plasma efficiently; Particularly the present invention is in order to enrichment blood platelet and obtain the platelet rich plasma of clinical treatment meaning from blood, seeks that a kind of operating process is simple, yield is high, PC is high in the blood plasma of enrichment method.It is hematoblastic concentrated and is separated that the present inventor finds to utilize gradient centrifugation to carry out, under specified operating conditions can to obtain with high yield containing the hematoblastic blood plasma of concentration height.
For this reason, first aspect present invention provides a kind of method extracting platelet rich plasma from blood, and it comprises the following steps:
A () makes the whole blood of collection be placed in container containing anti-coagulants, blood and anti-coagulants are fully mixed;
B () makes the blood being mixed with anti-coagulants be placed in centrifuge tube, it is centrifugal to carry out first time, makes blood substantially be divided into three layers;
C the superiors and most intermediate layer extract out and transfer in new centrifuge tube by (), mix, and it is centrifugal to carry out second time;
D () discards the blood plasma on centrifuge tube upper strata, utilize remaining blood plasma to make the blood platelet Eddy diffusion of precipitation, obtains platelet rich plasma (Platelet Rich Plasma:PRP).
Method according to a first aspect of the present invention, wherein whole blood described in step (a) is fresh whole blood.
Method according to a first aspect of the present invention, wherein whole blood described in step (a) is the whole blood of individual test subjects such as patient.Thus, the platelet rich plasma that the present invention obtains can be reused for this patient easily to treat relevant disease.
Method according to a first aspect of the present invention, wherein in step (a), anti-coagulants is selected from ethylenediamine tetra-acetic acid (Ethylene Diamine Tetraacetic Acid:EDTA), disodium ethylene diamine tetraacetate, calcio-disodium edetate, ACD (Acid Citrate Dextrose-Solution A:ACD-A) or its combination.
Method according to a first aspect of the present invention, wherein in step (a), the weight ratio of anti-coagulants and whole blood is 1:8 ~ 10, such as 1:9.
Method according to a first aspect of the present invention, wherein container described in step (a) can be centrifuge tube or blood taking bag etc.If centrifuge tube and this centrifuge tube capacity are applicable to directly centrifugal, then do not need in described step (b) to be separately placed in centrifuge tube, and can directly carry out centrifugal.
Method according to a first aspect of the present invention, wherein also be included in step (a) to get after making blood and anti-coagulants and (be such as less than 500ul in right amount, such as about 100ul) for measuring wherein hematoblastic quantity in blood, for use in follow-up process according to, monitoring.
Method is according to a first aspect of the present invention wherein reach the degree making blood substantially be divided into three layers in step (b) after first time is centrifugal.In one embodiment, wherein the superiors are containing hematoblastic plasma layer, and middle one deck is buffy coat (Buffy Coat layer, wherein containing blood platelet and leucocyte), the bottom be red blood cell layer.
Method according to a first aspect of the present invention, wherein centrifugal in step (b) rotating speed is 2000rpm ~ 3000rpm, such as 2200rpm ~ 2800rpm, such as 2300rpm ~ 2500rpm, such as about 2400rpm.
Method according to a first aspect of the present invention, wherein centrifugal in step (b) time is 1-10min, such as 2-8min, such as 3-5min, such as about 4min.
Method according to a first aspect of the present invention, wherein centrifugal in step (b) rotating speed is 2300rpm ~ 2500rpm, and the time is 3-5min.
Method according to a first aspect of the present invention, wherein centrifugal in step (b) rotating speed is 2400rpm, and the time is 4min.
Method according to a first aspect of the present invention, wherein in step (c), described most intermediate layer refer to as far as possible by whole for intermediate layer sucking-off but avoid extract red blood cell.
Method according to a first aspect of the present invention, wherein centrifugal in step (c) rotating speed is 1000rpm ~ 2000rpm, such as 1200rpm ~ 1800rpm, such as 1400rpm ~ 1600rpm, such as about 1500rpm.
Method according to a first aspect of the present invention, wherein centrifugal in step (c) time is 10-30min, such as 15-25min, such as 18-22min, such as about 20min.
Method according to a first aspect of the present invention, wherein centrifugal in step (c) rotating speed is 1400rpm ~ 1600rpm, and the time is 18-22min.
Method according to a first aspect of the present invention, wherein centrifugal in step (c) rotating speed is 1500rpm, and the time is 20min.
Method according to a first aspect of the present invention, wherein in step (c), gained upper strata and most of intermediate layer are after mixing, and wherein hematoblastic concentration is 1-5 times that is separated PC in front whole blood, such as 1.5 ~ 4 times, such as 2 ~ 3 times.
Method according to a first aspect of the present invention, wherein also be included in after the superiors of extraction and intermediate layer are mixed in step (c), get and (be such as less than 500ul in right amount, such as about 100ul) for measuring hematoblastic quantity wherein, contrasting for use in follow-up process, monitoring.
Method according to a first aspect of the present invention, wherein in step (d), described in discard centrifuge tube upper strata blood plasma refer to the blood plasma discarding centrifuge tube upper strata at least 2/4.In one embodiment, in step (d), described in discard centrifuge tube upper strata blood plasma refer to the blood plasma discarding centrifuge tube upper strata at least 3/4.Should illustrate at this, the blood platelet only containing low concentration in upper plasma.
Method according to a first aspect of the present invention, wherein in step (d), after discarding the blood plasma on centrifuge tube upper strata, can utilize that bottom is remaining (about 2/4, or preferred about 1/4) blood plasma makes the platelet suspension that precipitates again, can obtain platelet rich plasma of the present invention (PRP) thus.
Method according to a first aspect of the present invention, wherein in step (d) gained platelet rich plasma (PRP), hematoblastic concentration is 5-10 times that is separated PC in front whole blood, such as 6 ~ 8 times, such as 6 ~ 7 times.
Method according to a first aspect of the present invention, is wherein also included in step (d) after obtaining platelet rich plasma (PRP), gets appropriate (being such as less than 500ul, such as about 100ul) for measuring hematoblastic quantity wherein.
The thus obtained platelet rich plasma of the present invention (PRP) has the high feature of concentration, and recovery rate of blood platelet is high.
Second aspect present invention provides a kind of platelet rich plasma, wherein comprises 0.5 ~ 3 × 10
9the blood platelet of/ml concentration.Such as wherein comprise 1 ~ 2 × 10
9the blood platelet of/ml concentration.
Platelet rich plasma according to a second aspect of the present invention, it is obtained by the method described in the arbitrary embodiment of first aspect present invention substantially.
The present invention is further illustrated below.The document that the present invention quotes, and the document quoted in the document, their full content is incorporated to herein by reference.
In the present invention, in arbitrary technical scheme of either side of the present invention, its arbitrary technical characteristic is equally applicable to arbitrary embodiment of either side of the present invention, as long as they can not cause contradiction, and this being mutually useful in can do suitable amendment if desired.
In one embodiment of the invention, described whole blood is the whole blood of people.
In the present invention, a kind of method adopting specific Disposal Conditions to carry out platelet rich is provided.But it will be appreciated by those skilled in the art that and use Disposal Conditions of the present invention, utensil wherein used can do suitably variation, such as centrifugal device wherein.Such as (CN 102078644 A such as Wang Yue can be used, Chinese Patent Application No. 201110053979.5, denomination of invention: a kind of autologous platelet rich plasma extraction element of simple and effective and extracting method) in the used device be made up of syringe, intravenous transfusion device tube connector, venous detaining needle plastic cards and intravenous infusion apparatus needle, device described in the document may be used for the inventive method, and makes the inventive method be useful in the cleanliness factor keeping operating process.
Have been surprisingly found that, the present invention carries out hematoblastic concentrated with in the process be separated at research gradient centrifugation, obtains that a kind of operating process is simple, yield is high, PC is high in the blood plasma of enrichment method.
Detailed description of the invention
Can be conducted further description the present invention by the following examples, but scope of the present invention is not limited to following embodiment.One of skill in the art can understand, and under the prerequisite not deviating from the spirit and scope of the present invention, can carry out various change and modification to the present invention.The present invention carries out generality and/or concrete description to the material used in test and test method.Although for realizing many materials that the object of the invention uses and method of operating is well known in the art, the present invention still does description detailed as far as possible at this.
The following providing the general flow of method of operating of the present invention, as do not indicated in addition in Examples below, using the operating condition of general flow below:
(1) the collecting whole blood of patient to containing in the container of anti-coagulants, container can be centrifuge tube or blood taking bag.Anti-coagulants can use ethylenediamine tetra-acetic acid.The ratio of anti-coagulants and whole blood is 1:9.
(2) blood and anti-coagulants are fully mixed, avoid occurring blood coagulation.Extract 100 microlitres to calculate platelet counts.
(3) blood is assigned to centrifuge tube (15 milliliters).
(4) centrifuge tube is put into centrifuge, with rotating speed 2400rpm centrifugal 4 minutes.
(5) blood after centrifugal should be divided into three layers: the bottom be red blood cell, the superiors be that middle one deck is Buffy Coat, containing blood platelet and leucocyte containing thrombocyte plasma.
(6) utilize pipettor the blood plasma of the superiors and most buffy coat are extracted out and transfers in new centrifuge tube, avoid extracting red blood cell as far as possible.Extract 100 microlitres to calculate platelet counts.
(7) centrifuge tube containing blood plasma and buffy coat mixture is put into centrifuge, with rotating speed 1500rpm centrifugal 20 minutes.
(8) the low concentration thrombocyte plasma (Platelet Poor Plasma:PPP) of 3/4 is removed, utilize the remaining resuspended blood platelet precipitated of 1/4 blood plasma, obtain platelet rich plasma (PRP).
(9) 100 microlitre PRP are extracted to carry out platelet count.
embodiment 1: extract platelet rich plasma (PRP)
(1) PC in whole blood before being separated: 250 × 10
6/ ml, volume of whole blood is 10ml.Total platelet counts is 2.5 × 10
9.
(2) PC in first time centrifugal rear blood plasma: 570 × 10
6/ ml (concentrated 2.28 times), Plasma volumes is 4ml.Total platelet counts is 2.28 × 10
9.The rate of recovery is 91.2%.
(3) (PRP) PC in the centrifugal rear blood plasma of second time: 1685 × 10
6/ ml (concentrated 6.74 times), Plasma volumes is 1.2ml, obtains platelet rich plasma.Total platelet counts is 2.02 × 10
9.Ult rec is 80.8%.
embodiment 2: extract platelet rich plasma (PRP)
Use disodium ethylene diamine tetraacetate is anti-coagulants, and the weight ratio of anti-coagulants and whole blood is 1:8.
(1) PC in whole blood before being separated: 165 × 10
6/ ml, volume of whole blood is 10ml.Total platelet counts is 1.65 × 10
9.
(2) PC in first time centrifugal rear blood plasma: 444 × 10
6/ ml (concentrated 2.69 times), Plasma volumes is 3.5ml.Total platelet counts is 1.55 × 10
9.The rate of recovery is 93.9%.
(3) (PRP) PC in the centrifugal rear blood plasma of second time: 1105 × 10
6/ ml (concentrated 6.70 times), Plasma volumes is 1.2ml, obtains platelet rich plasma.Total platelet counts is 1.33 × 10
9.Ult rec is 80.6%.
embodiment 3: extract platelet rich plasma (PRP)
Use ACD is anti-coagulants.
(1) PC in whole blood before being separated: 210 × 10
6/ ml, volume of whole blood is 10ml.Total platelet counts is 2.1 × 10
9.
(2) PC in first time centrifugal rear blood plasma: 542 × 10
6/ ml (concentrated 2.58 times), Plasma volumes is 3.5ml.Total platelet counts is 1.90 × 10
9.The rate of recovery is 90.5%.
(3) (PRP) PC in the centrifugal rear blood plasma of second time: 1370 × 10
6/ ml (concentrated 6.52 times), Plasma volumes is 1.2ml, obtains platelet rich plasma.Total platelet counts is 1.64 × 10
9.Ult rec is 78.1%.
embodiment 4: extract platelet rich plasma (PRP)
The weight ratio of anti-coagulants and whole blood is 1:10, and first time centrifugally be 2300rpm and 5min, and second time is centrifugal is 1600rpm also 18 min.
(1) PC in whole blood before being separated: 195 × 10
6/ ml, volume of whole blood is 10ml.Total platelet counts is 1.95 × 10
9.
(2) PC in first time centrifugal rear blood plasma: 535 × 10
6/ ml (concentrated 2.74 times), Plasma volumes is 3.5ml.Total platelet counts is 1.87 × 10
9.The rate of recovery is 95.9%.
(3) (PRP) PC in the centrifugal rear blood plasma of second time: 1235 × 10
6/ ml (concentrated 6.33 times), Plasma volumes is 1.2ml, obtains platelet rich plasma.Total platelet counts is 1.48 × 10
9.Ult rec is 75.9%.
embodiment 5: extract platelet rich plasma (PRP)
First time centrifugally be 2500rpm and 3min, is centrifugally for the second time 1400rpm and 20min.
(1) PC in whole blood before being separated: 265 × 10
6/ ml, volume of whole blood is 10ml.Total platelet counts is 2.65 × 10
9.
(2) PC in first time centrifugal rear blood plasma: 612 × 10
6/ ml (concentrated 2.31 times), Plasma volumes is 4ml.Total platelet counts is 2.45 × 10
9.The rate of recovery is 92.5%.
(3) (PRP) PC in the centrifugal rear blood plasma of second time: 1839 × 10
6/ ml (concentrated 6.94 times), Plasma volumes is 1.2ml, obtains platelet rich plasma.Total platelet counts is 2.21 × 10
9.Ult rec is 83.4%.
Comparative example 1: except first time centrifugal rotational speed changes 2600rpm, 3000rpm, 3500rpm into, and outside these three kinds of rotating speeds all arrange 3min, 6min, 10min time, all the other operating conditions are all with embodiment 1.9 kinds of platelet rich plasmas that result display obtains like this, their blood platelet ult rec is only between 47 ~ 63%, and hematoblastic concentration is 1.8 ~ 3.7 times (can be described as final enrichment times in the present invention) of PC in whole blood before being separated.Such as 2600rpm 3min and 2600rpm in 6min two kinds of situations, blood platelet ult rec is respectively 56% and 61%, and final enrichment times is respectively 3.1 and 2.8.
Comparative example 2: except first time centrifugal rotational speed changes 2100rpm, 1700rpm, 1400rpm into, and outside these three kinds of rotating speeds all arrange 3min, 6min, 10min time, all the other operating conditions are all with embodiment 1.9 kinds of platelet rich plasmas that result display obtains like this, their blood platelet ult rec is only between 31 ~ 52%, and hematoblastic concentration is 1.3 ~ 2.5 times that are separated PC in front whole blood.Such as 2100rpm 3min and 2100rpm in 6min two kinds of situations, blood platelet ult rec is respectively 42% and 47%, and final enrichment times is respectively 1.9 and 2.2.
Comparative example 3: except second time centrifugal rotational speed changes 1800rpm, 2200rpm, 2600rpm into, and outside these three kinds of rotating speeds all arrange 18min, 20min, 25min time, all the other operating conditions are all with embodiment 1.9 kinds of platelet rich plasmas that result display obtains like this, their blood platelet ult rec is only between 52 ~ 63%, and hematoblastic concentration is 3.1 ~ 3.9 times that are separated PC in front whole blood.Such as 1800rpm 18min and 1800rpm in 20min two kinds of situations, blood platelet ult rec is respectively 53% and 57%, and final enrichment times is respectively 3.9 and 3.2.
Comparative example 4: except second time centrifugal rotational speed changes 1300rpm, 1000rpm, 700rpm into, and outside these three kinds of rotating speeds all arrange 18min, 20min, 25min time, all the other operating conditions are all with embodiment 1.9 kinds of platelet rich plasmas that result display obtains like this, their blood platelet ult rec is only between 33 ~ 46%, and hematoblastic concentration is 1.9 ~ 3.1 times that are separated PC in front whole blood.Such as 1300rpm 18min and 1300rpm in 20min two kinds of situations, blood platelet ult rec is respectively 43% and 35%, and final enrichment times is respectively 2.9 and 2.4.
Comparative example 5: first time is centrifugal changes 2300rpm into and 4min or 2500rpm 4min, and second time is centrifugal changes 1400rpm into and 20min or 1600rpm 20min, and all the other operating conditions are all with embodiment 1.4 kinds of platelet rich plasmas that result display obtains like this, their blood platelet ult rec is only between 48 ~ 61%, and hematoblastic concentration is 3.2 ~ 4.2 times that are separated PC in front whole blood.Such as first time is centrifugal be 2300rpm and 4min, second time centrifugal be 1600rpm 20min, blood platelet ult rec is 51%, and final enrichment times is respectively 3.7.
In addition, with reference to the operation of CN 102078644 A embodiment 1, blood platelet ult rec is 47%, and final enrichment times is respectively 3.45.
But the above result display of the present invention, the average platelet rate of recovery after the inventive method is centrifugal with second time in first time is respectively: 92.8% and 79.8%, and in the PRP after separation, hematoblastic concentration can reach the 6-7 of whole blood doubly.Show that the present invention's centrifugal time used and the setting of rotating speed can effectively be separated and reclaim the blood platelet in most of whole blood.
Claims (10)
1. from blood, extract the method for platelet rich plasma, it comprises the following steps:
A () makes the whole blood of collection be placed in container containing anti-coagulants, blood and anti-coagulants are fully mixed;
B () makes the blood being mixed with anti-coagulants be placed in centrifuge tube, it is centrifugal to carry out first time, makes blood substantially be divided into three layers;
C the superiors and most intermediate layer extract out and transfer in new centrifuge tube by (), mix, and it is centrifugal to carry out second time;
D () discards the blood plasma on centrifuge tube upper strata, utilize remaining blood plasma to make the blood platelet Eddy diffusion of precipitation, obtains platelet rich plasma (PlateletRich Plasma:PRP).
2. method according to claim 1, wherein in step (a):
Described whole blood is fresh whole blood; And/or
Described whole blood is the whole blood of individual test subjects such as patient.
3. according to the method for any one of claim 1 to 2, wherein in step (b):
Described anti-coagulants is selected from ethylenediamine tetra-acetic acid, disodium ethylene diamine tetraacetate, calcio-disodium edetate, ACD or its combination;
The weight ratio of described anti-coagulants and whole blood is 1:8 ~ 10; And/or
Also be included in described step (a) and get after making blood and anti-coagulants in right amount for measuring the step of hematoblastic quantity in wherein blood.
4. according to the method for any one of claims 1 to 3, wherein in step (b):
Reach the degree making blood substantially be divided into three layers after first time is centrifugal; And/or
Be reach the degree making blood substantially be divided into three layers after first time is centrifugal, wherein the superiors are containing hematoblastic plasma layer, and middle one deck is buffy coat, the bottom be red blood cell layer.
5. according to the method for any one of Claims 1-4, wherein in step (b):
Centrifugal rotating speed is 2000rpm ~ 3000rpm;
The centrifugal time is 1-10min; And/or
Centrifugal rotating speed is 2300rpm ~ 2500rpm, and the time is 3-5min.
6. according to the method for any one of claim 1 to 5, wherein in step (c):
Centrifugal rotating speed is 1000rpm ~ 2000rpm;
The centrifugal time is 10-30min;
Centrifugal rotating speed is 1400rpm ~ 1600rpm, and the time is 18-22min; And/or
Centrifugal rotating speed is 1500rpm, and the time is 20min.
7. according to the method for any one of claim 1 to 6, wherein in step (c):
Gained upper strata and most of intermediate layer are after mixing, and wherein hematoblastic concentration is 1-5 times that is separated PC in front whole blood; And/or
Also being included in after the superiors of extraction and intermediate layer are mixed, getting appropriate for measuring the step of hematoblastic quantity wherein.
8. according to the method for any one of claim 1 to 7, wherein in step (d):
The described blood plasma discarding centrifuge tube upper strata refers to the blood plasma discarding centrifuge tube upper strata at least 2/4;
The described blood plasma discarding centrifuge tube upper strata refers to the blood plasma discarding centrifuge tube upper strata at least 3/4;
After discarding the blood plasma on centrifuge tube upper strata, the blood plasma utilizing bottom to be left makes the platelet suspension precipitated again, obtains described platelet rich plasma;
In step (d) gained platelet rich plasma, hematoblastic concentration is 5-10 times that is separated PC in front whole blood; And/or
Also being included in step (d) after obtaining platelet rich plasma (PRP), getting appropriate for measuring the step of hematoblastic quantity wherein.
9. a platelet rich plasma, wherein comprises 0.5 ~ 3 × 10
9the blood platelet of/ml concentration.
10. platelet rich plasma according to claim 9, it is obtained by method described in any one of claim 1-8 substantially.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0233112A1 (en) * | 1986-01-24 | 1987-08-19 | TERUMO KABUSHIKI KAISHA trading as TERUMO CORPORATION | Method for separation of blood components and apparatus therefor |
| CN1261815A (en) * | 1997-07-01 | 2000-08-02 | 巴克斯特国际有限公司 | Systems and method for collecting diluted mononuclear cells |
| CN1300233A (en) * | 1999-04-12 | 2001-06-20 | 丰收技术股份有限公司 | Method and apparatus for producing platelet rich plasma and/or platelet |
| CN1583213A (en) * | 2004-06-11 | 2005-02-23 | 西安瑞捷生物医疗技术有限公司 | Apparatus for preparing thrombocyte enriched blood plasma and preparing method thereof |
| EP1252918B1 (en) * | 1992-10-22 | 2005-03-02 | Baxter International Inc. | Whole blood centrifugal separation apparatus and method |
| JP2008050688A (en) * | 2006-07-11 | 2008-03-06 | Senju Metal Ind Co Ltd | Sliding material and method for its manufacture |
| CN101528243A (en) * | 2006-10-27 | 2009-09-09 | 尼普洛株式会社 | Platelet-rich plasma separator and platelet-rich plasma separation method |
| TW201031417A (en) * | 2009-02-26 | 2010-09-01 | Win-Ping Deng | Platelet-rich plasma (PRP) derived growth factor complex, method of producing the same, medical compound comprising the same, and method for enhancing growth of tissue of patient |
| CN102078644A (en) * | 2011-03-08 | 2011-06-01 | 吉林大学 | Simple, convenient and efficient device and method for extracting platelet-rich plasma |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1383831A (en) * | 2001-04-28 | 2002-12-11 | 益阳市中心血站 | Method and device for preparing concentrated thrombocyte |
| KR101087653B1 (en) * | 2008-11-13 | 2011-11-30 | 이희영 | Preparing for prp |
-
2012
- 2012-07-30 CN CN201410570704.2A patent/CN104383726B/en active Active
- 2012-07-30 CN CN201210267055.XA patent/CN102755770B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0233112A1 (en) * | 1986-01-24 | 1987-08-19 | TERUMO KABUSHIKI KAISHA trading as TERUMO CORPORATION | Method for separation of blood components and apparatus therefor |
| EP1252918B1 (en) * | 1992-10-22 | 2005-03-02 | Baxter International Inc. | Whole blood centrifugal separation apparatus and method |
| CN1261815A (en) * | 1997-07-01 | 2000-08-02 | 巴克斯特国际有限公司 | Systems and method for collecting diluted mononuclear cells |
| CN1300233A (en) * | 1999-04-12 | 2001-06-20 | 丰收技术股份有限公司 | Method and apparatus for producing platelet rich plasma and/or platelet |
| CN1583213A (en) * | 2004-06-11 | 2005-02-23 | 西安瑞捷生物医疗技术有限公司 | Apparatus for preparing thrombocyte enriched blood plasma and preparing method thereof |
| JP2008050688A (en) * | 2006-07-11 | 2008-03-06 | Senju Metal Ind Co Ltd | Sliding material and method for its manufacture |
| CN101528243A (en) * | 2006-10-27 | 2009-09-09 | 尼普洛株式会社 | Platelet-rich plasma separator and platelet-rich plasma separation method |
| TW201031417A (en) * | 2009-02-26 | 2010-09-01 | Win-Ping Deng | Platelet-rich plasma (PRP) derived growth factor complex, method of producing the same, medical compound comprising the same, and method for enhancing growth of tissue of patient |
| CN102078644A (en) * | 2011-03-08 | 2011-06-01 | 吉林大学 | Simple, convenient and efficient device and method for extracting platelet-rich plasma |
Non-Patent Citations (1)
| Title |
|---|
| 李明，张长青，袁霆，陈圣宝等: "富血小板血浆制备套装的评估研究", 《中国修复重建外壳杂志》, vol. 25, no. 1, 10 December 2010 (2010-12-10) * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105107233A (en) * | 2015-07-24 | 2015-12-02 | 上海市第六人民医院 | Preparation method and device for leukocyte-depleted platelet rich plasma |
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| CN106727700A (en) * | 2016-11-29 | 2017-05-31 | 第五空间健康管理江苏有限公司 | Prepare the method for platelet rich plasma PRP and the purposes of the platelet rich plasma |
| CN106701674A (en) * | 2016-12-29 | 2017-05-24 | 上海新肌生物科技有限公司 | Purification and extraction process of resuscitation-promoting factors capable of increasing survival rate of transplanted adipocytes or filled adipocytes |
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| CN109692761A (en) * | 2018-12-25 | 2019-04-30 | 山东博森医学工程技术有限公司 | A kind of platelet rich plasma extraction element and its method |
| CN112237755A (en) * | 2019-07-18 | 2021-01-19 | 北京纳通医学科技研究院有限公司 | Preparation method and preparation device of platelet rich plasma and prepared platelet rich plasma |
| CN112237755B (en) * | 2019-07-18 | 2023-12-05 | 北京纳通医学科技研究院有限公司 | Preparation method and preparation device of platelet-rich plasma and prepared platelet-rich plasma |
| CN110538196A (en) * | 2019-08-13 | 2019-12-06 | 杭州易文赛生物技术有限公司 | Platelet-rich plasma and method for extracting platelet-rich plasma |
| CN113488180A (en) * | 2021-07-28 | 2021-10-08 | 中国医学科学院医学信息研究所 | Clinical guideline knowledge modeling method and system |
| CN113488180B (en) * | 2021-07-28 | 2023-07-18 | 中国医学科学院医学信息研究所 | Clinical guideline knowledge modeling method and system |
| CN115633679A (en) * | 2022-10-24 | 2023-01-24 | 复旦大学附属中山医院 | Separation and preservation solution and separation method of platelet-rich plasma |
| CN115633679B (en) * | 2022-10-24 | 2023-10-31 | 江苏美赛尔生物科技有限公司 | Separation preservation solution and separation method for platelet-rich plasma |
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| CN104383726B (en) | 2016-05-25 |
| CN102755770A (en) | 2012-10-31 |
| CN102755770B (en) | 2014-12-03 |
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