CN102755770B - Extraction method of platelet rich plasma (PRP) and extracted PRP - Google Patents

Extraction method of platelet rich plasma (PRP) and extracted PRP Download PDF

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CN102755770B
CN102755770B CN201210267055.XA CN201210267055A CN102755770B CN 102755770 B CN102755770 B CN 102755770B CN 201210267055 A CN201210267055 A CN 201210267055A CN 102755770 B CN102755770 B CN 102755770B
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blood
step
plasma
according
platelet
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CN201210267055.XA
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CN102755770A (en
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林卓衡
陈俊峯
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博雅干细胞科技有限公司
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Abstract

The invention relates to an extraction method of platelet rich plasma (PRP) and extracted PRP and particularly relates to a method for extracting PRP from blood. The method comprises the following steps of: (a) putting the collected whole blood in a container containing an anticoagulant, and fully and uniformly mixing the blood and the anticoagulant; (b) putting the blood mixed with the anticoagulant in a centrifuge tube, carrying out primary centrifugation, and enabling the blood to be divided into three layers; (c) extracting the uppermost layer and most of the middle layer, transferring the uppermost layer and most of the middle layer into a new centrifuge tube, mixing uniformly and carrying out secondary centrifugation; and (d) discarding the plasma at the upper layer of the centrifuge tube, and resuspending the precipitated plasma by utilizing the remaining plasma, thus obtaining the PRP. The method provided by the invention has the advantages that the operation process is simple, the yield is high and the concentration of platelet in rich plasma is high.

Description

The extracting method of platelet rich plasma and the platelet rich plasma of extraction

Technical field

The invention belongs to clinical medicine technical field, be specifically related to a kind of extraction and be rich in the method for hematoblastic blood plasma, the platelet rich plasma that the inventive method obtains under given conditions has advantages of that PC is high, high at least one aspect of recovery rate of blood platelet.

Background technology

Be rich in hematoblastic blood plasma, can be described as in the present invention platelet rich plasma (Platelet Rich Plasma, PRP), it is the blood platelet blood plasma containing high concentration obtaining by centrifugal blood.The blood platelet of human body be except providing cohesion in hemostasis, also contains much and wound healing and the bone relevant multiple growth factor of regenerating 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, Differentiation and proliferation to cell plays facilitation, these growth factors have cooperative response each other, factor interaction and impact with other promotion cytoactives, jointly maintaining 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 is processed, can be used for the treatment of relevant disease, thereby source is abundant, draws materials conveniently, preparation method is simple and can allow health absorb.Self PRP uses and can avoid virus to propagate and the problem appearance of immunological rejection.Therefore PRP is widely applied in different medical domains.As orthopaedics, the department of stomatology, decorative sursery, sports medical science and aesthetic medicine.

PRP goes back the preparation method that neither one is unified so far, the preparation principle of PRP is mainly to utilize various composition sinking speed differences in blood, by centrifugal by blood layering.Thereby obtain containing hematoblastic blood plasma, and utilize centrifugal principle further concentrated to obtain the blood platelet blood plasma of high concentration.In human whole blood, hematoblastic concentration is generally 1 ~ 3 × 105/ml.Research thinks that the PRP PC after concentrating should be 3-4 times of whole blood PC.Research also finds that the raising of PC in PRP can effectively improve propagation and the differentiation capability of stem cell and can significantly increase the expression of fibroblastic propagation and type i collagen albumen.

Therefore effectively separate 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) a kind of method for extracting plastocyte of mouse disclosed, mouse socket of the eye venous blood sampling, be placed in anti-freezing centrifuge tube, centrifugal after standing 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, and abandoning supernatant, pipe end sediment is blood platelet.Be 1% ammonium oxalate solution to dripping 50~100 Μ L mass concentrations in test tube, stir gently with glass bar, then to add mass concentration be 1% ammonium oxalate solution 2~3ML, leave standstill 5MIN, make erythrocytolysis.The centrifugal 10MIN of 3500RPM, supernatant discarded, adds blood platelet cleaning solution piping and druming repeatedly, makes to become thrombocyte suspension, the centrifugal 10MIN of 3300RPM, supernatant discarded, adds a small amount of blood platelet cleaning solution and repeatedly blows and beats and make it to become suspension.This blood platelet will be served as after antigen immune cavy, 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 well the not good problem of blood platelet purity.Although this method of carrying out blood platelet enrichment for mouse blood it is believed that can obtain concentration reaches 10 10blood platelet, but unexposed its hematoblastic yield of the method, and method is more numerous and diverse in operation, is difficult to be applicable to the processing of human blood.

In addition, (the CN102078644A such as Wang Yue, Chinese Patent Application No. 201110053979.5, denomination of invention: a kind of autologous platelet rich plasma extraction element and extracting method of simple and effective) in a kind of autologous platelet rich plasma extraction element and extracting method of simple and effective are disclosed, in the autologous platelet rich plasma extracting, contain multiple composite growth factor, can effectively promote the reparation of tissue.Formed by 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, and intravenous infusion apparatus needle needs the autologous platelet rich plasma after position injection is extracted in order to skin heart puncture, extraction anti-coagulants and activator and 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 being useful aspect the operability of lifting blood platelet extraction.

Therefore obtain the platelet rich plasma of clinical treatment meaning for enrichment blood platelet from blood, seek that a kind of operating process is simple, yield is high, the high method of PC in the blood plasma of enrichment, being that those skilled in the art are urgent expects.

Summary of the invention

The object of the invention is to explore one simply with separate efficiently and concentrated blood plasma in hematoblastic method; Particularly the present invention obtains the platelet rich plasma of clinical treatment meaning for enrichment blood platelet from blood, seeks that a kind of operating process is simple, yield is high, the high method of PC in the blood plasma of enrichment.It is hematoblastic concentrated and separates that the inventor finds to utilize gradient centrifugation to carry out, can be to obtain with high yield containing the hematoblastic blood plasma of concentration height under specific operation condition.

For this reason, first aspect present invention provides a kind of method of extracting platelet rich plasma from blood, and it comprises the following steps:

(a) make the whole blood of collection be placed in the container that contains anti-coagulants, blood and anti-coagulants are fully mixed;

(b) blood that makes to be mixed with anti-coagulants is placed in centrifuge tube, carries out centrifugally for the first time, makes blood substantially be divided into three layers;

(c) the superiors and most intermediate layer are extracted out and transferred in new centrifuge tube, mix, carry out centrifugal for the second time;

(d) discard the blood plasma on centrifuge tube upper strata, utilize remaining blood plasma to make the blood platelet Eddy diffusion of precipitation, obtain platelet rich plasma (Platelet Rich Plasma:PRP).

According to the method for first aspect present invention, wherein described in step (a), whole blood is fresh whole blood.

According to the method for first aspect present invention, wherein described in step (a), whole blood is for example patient's of tested individuality whole blood.Thus, the platelet rich plasma that the present invention obtains can be reused for this patient easily with treatment relevant disease.

According to the method for first aspect 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.

According to the method for first aspect present invention, wherein in step (a), the weight ratio of anti-coagulants and whole blood is 1:8 ~ 10, for example 1:9.

According to the method for first aspect present invention, wherein described in step (a), container can be centrifuge tube or blood taking bag etc.Directly centrifugal if centrifuge tube and this centrifuge tube capacity are applicable to, in described step (b), do not need to be separately placed in centrifuge tube, and can directly carry out centrifugal.

According to the method for first aspect present invention, wherein in step (a), be also included in and make to get in right amount and (be for example less than 500ul after blood and anti-coagulants, for example about 100ul) for measuring the wherein hematoblastic quantity of blood, contrast, monitor for use in follow-up process.

According to the method for first aspect present invention, in step (b), after centrifugal for the first time, be wherein to reach to make blood substantially be divided into the degree of three layers.In one embodiment, wherein the superiors are containing hematoblastic plasma layer, and middle one deck is buffy coat (Buffy Coat layer, wherein contains blood platelet and leucocyte), the bottom be red blood cell layer.

According to the method for first aspect present invention, wherein in step (b), centrifugal rotating speed is 2000rpm ~ 3000rpm, for example 2200rpm ~ 2800rpm, for example 2300rpm ~ 2500rpm, for example about 2400rpm.

According to the method for first aspect present invention, wherein in step (b), the centrifugal time is 1-10min, for example 2-8min, for example 3-5min, for example about 4min.

According to the method for first aspect present invention, wherein in step (b), centrifugal rotating speed is 2300rpm ~ 2500rpm, and the time is 3-5min.

According to the method for first aspect present invention, wherein in step (b), centrifugal rotating speed is 2400rpm, and the time is 4min.

According to the method for first aspect present invention, wherein in step (c), described most intermediate layer refers to as far as possible whole intermediate layer sucking-offs but avoids extracting red blood cell.

According to the method for first aspect present invention, wherein in step (c), centrifugal rotating speed is 1000rpm ~ 2000rpm, for example 1200rpm ~ 1800rpm, for example 1400rpm ~ 1600rpm, for example about 1500rpm.

According to the method for first aspect present invention, wherein in step (c), the centrifugal time is 10-30min, for example 15-25min, for example 18-22min, for example about 20min.

According to the method for first aspect present invention, wherein in step (c), centrifugal rotating speed is 1400rpm ~ 1600rpm, and the time is 18-22min.

According to the method for first aspect present invention, wherein in step (c), centrifugal rotating speed is 1500rpm, and the time is 20min.

According to the method for first aspect present invention, wherein in step (c) gained upper strata and most of intermediate layer after mixing, wherein hematoblastic concentration be before separating in whole blood the 1-5 of PC doubly, for example 1.5 ~ 4 times, for example 2 ~ 3 times.

According to the method for first aspect present invention, wherein in step (c), be also included in and make the superiors that extract and after intermediate layer mixes, get in right amount and (be for example less than 500ul, for example about 100ul) for measuring hematoblastic quantity wherein, contrast, monitor for use in follow-up process.

According to the method for first aspect present invention, wherein in step (d), described in discard centrifuge tube upper strata blood plasma refer to the blood plasma that discards 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 that discards centrifuge tube upper strata at least 3/4.Should illustrate at this, in upper plasma, only contain the blood platelet of low concentration.

According to the method for first aspect present invention, wherein in step (d), discard after the blood plasma on centrifuge tube upper strata, can utilize bottom remaining (approximately 2/4, or preferred approximately 1/4) blood plasma makes the platelet suspension precipitating again, can obtain thus platelet rich plasma of the present invention (PRP).

According to the method for first aspect present invention, wherein in step (d) gained platelet rich plasma (PRP) hematoblastic concentration be before separating in whole blood the 5-10 of PC doubly, for example 6 ~ 8 times, for example 6 ~ 7 times.

According to the method for first aspect present invention, wherein in step (d), be also included in and obtain after platelet rich plasma (PRP), get appropriate (being for example less than 500ul, for example about 100ul) for measuring hematoblastic quantity wherein.

The thus obtained platelet rich plasma of the present invention (PRP) has the advantages that concentration is high, 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.For example wherein comprise 1 ~ 2 × 10 9the blood platelet of/ml concentration.

According to the platelet rich plasma of second aspect present invention, it is to be 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 of quoting 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 characterictic 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 be done suitable amendment if desired.

In one embodiment of the invention, described whole blood is people's whole blood.

In the present invention, provide a kind of method that adopts specific Disposal Conditions to carry out blood platelet enrichment.But it will be appreciated by those skilled in the art that and use Disposal Conditions of the present invention, wherein utensil used can be done suitably variation, for example centrifugal device wherein.For example can use (the CN102078644A such as Wang Yue, Chinese Patent Application No. 201110053979.5, denomination of invention: a kind of autologous platelet rich plasma extraction element and extracting method of simple and effective) in the used device being formed by syringe, intravenous transfusion device tube connector, venous detaining needle plastic cards and intravenous infusion apparatus needle, the device that the document is recorded can be for the inventive method, and to make the inventive method be useful in the cleanliness factor that keeps operating process.

Have been surprisingly found that, the present invention carries out hematoblastic concentrated with in the process separating at research gradient centrifugation, has obtained that a kind of operating process is simple, yield is high, the high method of PC in the blood plasma of enrichment.

Detailed description of the invention

Can conduct 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 is not deviating under the prerequisite of the spirit and scope of the present invention, can carry out various variations and modification to the present invention.The present invention carries out generality and/or concrete description to the material and the test method that use in test.Although be well known in the art for realizing many materials and the method for operating that the object of the invention uses, the present invention still does detailed as far as possible description at this.

The general flow of method of operating of the present invention is below provided, in embodiment below as, has not indicated in addition, used the operating condition of general flow below:

(1) patient's whole blood is collected in the container that contains anti-coagulants to container it is 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, contains blood platelet and leucocyte containing blood platelet blood plasma.

(6) utilize pipettor that the blood plasma of the superiors and most buffy coat are extracted out and transferred in new centrifuge tube, avoid extracting red blood cell as far as possible.Extract 100 microlitres to calculate platelet counts.

(7) centrifuge tube that contains blood plasma and buffy coat mixture is put into centrifuge, with rotating speed 1500rpm centrifugal 20 minutes.

(8) 3/4 low concentration blood platelet blood plasma (Platelet Poor Plasma:PPP) is removed, utilize the remaining resuspended blood platelet precipitating of 1/4 blood plasma, obtain platelet rich plasma (PRP).

(9) extract 100 microlitre PRP to carry out platelet count.

embodiment 1: extract platelet rich plasma (PRP)

(1) separate PC in front whole blood: 250 × 10 6/ ml, volume of whole blood is 10ml.Total platelet counts is 2.5 × 10 9.

(2) PC in centrifugal rear blood plasma for the first time: 570 × 10 6/ ml (concentrated 2.28 times), blood plasma volume is 4ml.Total platelet counts is 2.28 × 10 9.The rate of recovery is 91.2%.

(3) (PRP) PC in centrifugal rear blood plasma for the second time: 1685 × 10 6/ ml (concentrated 6.74 times), blood plasma volume 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) separate PC in front whole blood: 165 × 10 6/ ml, volume of whole blood is 10ml.Total platelet counts is 1.65 × 10 9.

(2) PC in centrifugal rear blood plasma for the first time: 444 × 10 6/ ml (concentrated 2.69 times), blood plasma volume is 3.5ml.Total platelet counts is 1.55 × 10 9.The rate of recovery is 93.9%.

(3) (PRP) PC in centrifugal rear blood plasma for the second time: 1105 × 10 6/ ml (concentrated 6.70 times), blood plasma volume 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) separate PC in front whole blood: 210 × 10 6/ ml, volume of whole blood is 10ml.Total platelet counts is 2.1 × 10 9.

(2) PC in centrifugal rear blood plasma for the first time: 542 × 10 6/ ml (concentrated 2.58 times), blood plasma volume is 3.5ml.Total platelet counts is 1.90 × 10 9.The rate of recovery is 90.5%.

(3) (PRP) PC in centrifugal rear blood plasma for the second time: 1370 × 10 6/ ml (concentrated 6.52 times), blood plasma volume 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 centrifugal is for the first time 2300rpm 5min, and centrifugal is for the second time 1600rpm 18min.

(1) separate PC in front whole blood: 195 × 10 6/ ml, volume of whole blood is 10ml.Total platelet counts is 1.95 × 10 9.

(2) PC in centrifugal rear blood plasma for the first time: 535 × 10 6/ ml (concentrated 2.74 times), blood plasma volume is 3.5ml.Total platelet counts is 1.87 × 10 9.The rate of recovery is 95.9%.

(3) (PRP) PC in centrifugal rear blood plasma for the second time: 1235 × 10 6/ ml (concentrated 6.33 times), blood plasma volume 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)

Centrifugal is for the first time 2500rpm 3min, and centrifugal is for the second time 1400rpm 20min.

(1) separate PC in front whole blood: 265 × 10 6/ ml, volume of whole blood is 10ml.Total platelet counts is 2.65 × 10 9.

(2) PC in centrifugal rear blood plasma for the first time: 612 × 10 6/ ml (concentrated 2.31 times), blood plasma volume is 4ml.Total platelet counts is 2.45 × 10 9.The rate of recovery is 92.5%.

(3) (PRP) PC in centrifugal rear blood plasma for the second time: 1839 × 10 6/ ml (concentrated 6.94 times), blood plasma volume is 1.2ml, obtains platelet rich plasma.Total platelet counts is 2.21 × 10 9.Ult rec is 83.4%.

Comparative example 1: except centrifugal rotational speed for the first time changes 2600rpm, 3000rpm, 3500rpm into, and these three kinds of rotating speeds all arrange 3min, 6min, outside the 10min time, all the other operating conditions are all with embodiment 1.Result shows 9 kinds of platelet rich plasmas that obtain like this, their blood platelet ult rec is only between 47 ~ 63%, and hematoblastic concentration is before separating in whole blood 1.8 ~ 3.7 times (can be described as in the present invention final enrichment times) of PC.For example, in two kinds of situations of 2600rpm 3min and 2600rpm 6min, blood platelet ult rec is respectively 56% and 61%, and final enrichment times is respectively 3.1 and 2.8.

Comparative example 2: except centrifugal rotational speed for the first time changes 2100rpm, 1700rpm, 1400rpm into, and these three kinds of rotating speeds all arrange 3min, 6min, outside the 10min time, all the other operating conditions are all with embodiment 1.Result shows 9 kinds of platelet rich plasmas that obtain like this, and their blood platelet ult rec is only between 31 ~ 52%, and hematoblastic concentration is before separating in whole blood 1.3 ~ 2.5 times of PC.For example, in two kinds of situations of 2100rpm 3min and 2100rpm 6min, blood platelet ult rec is respectively 42% and 47%, and final enrichment times is respectively 1.9 and 2.2.

Comparative example 3: except centrifugal rotational speed for the second time changes 1800rpm, 2200rpm, 2600rpm into, and these three kinds of rotating speeds all arrange 18min, 20min, outside the 25min time, all the other operating conditions are all with embodiment 1.Result shows 9 kinds of platelet rich plasmas that obtain like this, and their blood platelet ult rec is only between 52 ~ 63%, and hematoblastic concentration is before separating in whole blood 3.1 ~ 3.9 times of PC.For example, in two kinds of situations of 1800rpm 18min and 1800rpm 20min, blood platelet ult rec is respectively 53% and 57%, and final enrichment times is respectively 3.9 and 3.2.

Comparative example 4: except centrifugal rotational speed for the second time changes 1300rpm, 1000rpm, 700rpm into, and these three kinds of rotating speeds all arrange 18min, 20min, outside the 25min time, all the other operating conditions are all with embodiment 1.Result shows 9 kinds of platelet rich plasmas that obtain like this, and their blood platelet ult rec is only between 33 ~ 46%, and hematoblastic concentration is before separating in whole blood 1.9 ~ 3.1 times of PC.For example, in two kinds of situations of 1300rpm 18min and 1300rpm 20min, blood platelet ult rec is respectively 43% and 35%, and final enrichment times is respectively 2.9 and 2.4.

Comparative example 5: centrifugally for the first time change 2300rpm 4min or 2500rpm 4min into, and centrifugal 1400rpm 20min or 1600rpm the 20min of changing into for the second time, all the other operating conditions are all with embodiment 1.Result shows 4 kinds of platelet rich plasmas that obtain like this, and their blood platelet ult rec is only between 48 ~ 61%, and hematoblastic concentration is before separating in whole blood 3.2 ~ 4.2 times of PC.For example for the first time centrifugal be 2300rpm 4min, centrifugal be for the second time 1600rpm 20min, blood platelet ult rec is 51%, final enrichment times is respectively 3.7.

In addition, with reference to the operation of CN 102078644A embodiment 1, blood platelet ult rec is 47%, and final enrichment times is respectively 3.45.

But the above result of the present invention shows, the average platelet rate of recovery of the inventive method after for the first time and for the second time centrifugal is respectively: 92.8% and 79.8%, and the 6-7 that in the PRP after separation, hematoblastic concentration can reach whole blood is doubly.Show that centrifugal time that the present invention is used and the setting of rotating speed can effectively separate and reclaim the blood platelet in most of whole blood.

Claims (15)

1. the method for extracting platelet rich plasma from blood, it comprises the following steps:
(a) make the whole blood of collection be placed in the container that contains anti-coagulants, blood and anti-coagulants are fully mixed;
(b) blood that makes to be mixed with anti-coagulants is placed in centrifuge tube, and carrying out centrifugal for the first time, centrifugal rotating speed is 2400 rpm, and the time is 4min, makes blood substantially be divided into three layers;
(c) the superiors and most intermediate layer are extracted out and transferred in new centrifuge tube, mix, carrying out centrifugal for the second time, centrifugal rotating speed is 1500 rpm, and the time is 20min;
(d) discard the blood plasma on centrifuge tube upper strata, utilize remaining blood plasma to make the blood platelet Eddy diffusion of precipitation, obtain platelet rich plasma.
2. according to the process of claim 1 wherein that described in step (a), whole blood is fresh whole blood.
3. according to the process of claim 1 wherein that described in step (a), whole blood is the whole blood of tested individuality.
4. according to the process of claim 1 wherein that described in step (a), anti-coagulants is selected from ethylenediamine tetra-acetic acid, disodium ethylene diamine tetraacetate, calcio-disodium edetate, ACD or its combination.
5. according to the process of claim 1 wherein that the weight ratio of anti-coagulants described in step (a) and whole blood is 1:8 ~ 10.
According to the process of claim 1 wherein be also included in step (a) get after blood and anti-coagulants are mixed appropriate for measuring the wherein step of the hematoblastic quantity of blood.
7. according to the process of claim 1 wherein in step (b), after centrifugal for the first time, be to reach to make blood substantially be divided into the degree of three layers.
8. according to the method for claim 1, wherein in step (b), after centrifugal for the first time, be to reach to make blood substantially be divided into the degree of three layers, wherein the superiors are containing hematoblastic plasma layer, middle one deck is buffy coat, the bottom be red blood cell layer.
9. according to the process of claim 1 wherein in step (c): gained upper strata and most of intermediate layer are after mixing, and wherein hematoblastic concentration is that before separating, in whole blood, the 2-3 of PC is doubly.
10. according to the process of claim 1 wherein in step (c), after being also included in the superiors that extract and intermediate layer being mixed, get appropriate for measuring the step of hematoblastic quantity wherein.
11. according to the process of claim 1 wherein in step (d), described in discard centrifuge tube upper strata blood plasma refer to the blood plasma that discards centrifuge tube upper strata at least 2/4.
12. according to the process of claim 1 wherein in step (d), described in discard centrifuge tube upper strata blood plasma refer to the blood plasma that discards centrifuge tube upper strata at least 3/4.
13. according to the process of claim 1 wherein in step (d), discards after the blood plasma on centrifuge tube upper strata, utilizes the remaining blood plasma in bottom again to make the platelet suspension precipitating, and obtains described platelet rich plasma.
14. according to the process of claim 1 wherein in step (d), and in step (d) gained platelet rich plasma, hematoblastic concentration is that before separating, in whole blood, the 5-10 of PC is doubly.
15. according to the process of claim 1 wherein in step (d), is also included in and obtains after platelet rich plasma in step (d), gets appropriate for measuring the step of hematoblastic quantity wherein.
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