WO2021098011A1 - 一种自体白细胞和血小板血浆综合提取方法 - Google Patents

一种自体白细胞和血小板血浆综合提取方法 Download PDF

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WO2021098011A1
WO2021098011A1 PCT/CN2019/129381 CN2019129381W WO2021098011A1 WO 2021098011 A1 WO2021098011 A1 WO 2021098011A1 CN 2019129381 W CN2019129381 W CN 2019129381W WO 2021098011 A1 WO2021098011 A1 WO 2021098011A1
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翟雯君
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四川我爱我来汽车俱乐部有限公司
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    • A61K35/14—Blood; Artificial blood
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants

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  • the invention relates to the technical field of plasma extraction, in particular to a method for comprehensively extracting autologous leukocytes and platelet plasma.
  • Platelet Rich Plasma is high-concentration platelet-containing plasma obtained by centrifuging blood.
  • Human platelets can produce proteins with cell adhesion function (fibrin, fibronectin & vitronectin) at high concentrations, Chinese names: fibronectin, fibronectin, and when the pH value drops from 7.0 to 7.2 to 6.5 to 6.7, Adding appropriate calcium chloride can promote the platelet secretion of 9 growth factors that promote wound, tissue healing and cell regeneration, so PRP is also called "growth factor-rich plasma”.
  • PRP can quickly stop bleeding, relieve pain, and accelerate wound healing, which can greatly reduce the formation of postoperative scars.
  • PRP is extracted from the patient's own blood, after being extracted and enriched, it can be used for the treatment of related diseases, so it has abundant sources and convenient materials that can be absorbed by the body.
  • the use of own PRP can avoid the emergence of problems such as virus transmission and immune rejection.
  • the present invention provides a comprehensive extraction method for autologous leukocytes and platelet plasma, which is simple to operate, can quickly extract leukocytes and platelet plasma, does not add anticoagulants and other additives, and avoids users after extraction. In case of allergies, pain and other problems, it can be injected directly, which is convenient for clinical use and improves the therapeutic effect.
  • the technical solution adopted by the present invention to solve its technical problems is to provide a comprehensive extraction method for autologous leukocytes and platelet plasma, which includes the following steps:
  • step (1) Put the short-cut syringe in step (1) into a sterile centrifugal sleeve, and centrifuge at 1-20°C for 2-6min;
  • step (3) The blood in the syringe after centrifugation in step (2) is divided into two layers, and the upper layer of plasma is extracted through the medical three-way tube using another syringe to obtain leukocyte and platelet plasma.
  • step (1) when cutting the syringe core rod in step (1), cut off the excess core rod in the recess behind the piston.
  • the centrifugal force is 70-1000 g.
  • the plasma concentration obtained in step (3) is 1 to 2 times white blood cells and 1 to 3 times platelet plasma.
  • the blood does not need to add additives such as anticoagulants during the preparation process.
  • the present invention has the following advantages:
  • the extraction method provided by the present invention uses an ice bag to wrap a conventional syringe during blood collection to immediately lower the blood temperature to 1-20°C. At this temperature, the enzyme activity is reduced, the blood is not easy to coagulate, and leukocytes and platelet plasma can be quickly extracted ; No additives such as anticoagulants are added to avoid allergies, pain and other problems of the user after extraction; it can be injected directly, which is convenient for clinical use and improves the treatment effect; the upper plasma after centrifugation is liquid plasma containing white blood cells and platelets, which is injected After reaching the human body, a gel is formed at the injection site, and the effect is maintained for 7 to 10 days. It can be widely used in medical cosmetology, hair growth, osteoarthritis, sports injuries, tooth anti-aging and other medical fields.
  • the syringes are all disposable sterile consumables, and the centrifuge tube is sterilized and sealed to prevent cross-infection and simplify the operation steps.
  • the plasma extracted by this program does not require an activator when used, and the extraction time is reduced by 90% compared with conventional methods. It is convenient for direct injection and has higher clinical safety.
  • the extract is completely derived from autologous plasma, and its components are extracted by previous methods. The ingredients are the same, and because there are no additives, there will be no risks of allergies, rejection and infection, and it is safer and more reliable.
  • a comprehensive extraction method for autologous leukocytes and platelet plasma includes the following steps:
  • step (1) Put the short-cut syringe in step (1) into a sterile centrifugal sleeve, and centrifuge for 2 minutes at a temperature of 1-20°C with a centrifugal force of 70g;
  • step (3) The blood in the syringe after centrifugation in step (2) is divided into two layers, and the upper layer of plasma is extracted through a medical three-way tube using another syringe to obtain 1 to 2 times white blood cells and 1 to 3 times platelet plasma.
  • a comprehensive extraction method for autologous leukocytes and platelet plasma includes the following steps:
  • step (1) Put the short-cut syringe in step (1) into a sterile centrifugal sleeve, and centrifuge at 1-20°C for 4 minutes, with a centrifugal force of 500g;
  • step (3) The blood in the syringe after centrifugation in step (2) is divided into two layers, and the upper layer of plasma is extracted through a medical three-way tube using another syringe to obtain 1 to 2 times white blood cells and 1 to 3 times platelet plasma.
  • a comprehensive extraction method for autologous leukocytes and platelet plasma includes the following steps:
  • step (1) Put the short-cut syringe in step (1) into a sterile centrifugal sleeve, and centrifuge for 6 min at a temperature of 1-20°C with a centrifugal force of 1000 g;
  • step (3) The blood in the syringe after centrifugation in step (2) is divided into two layers, and the upper layer of plasma is extracted through a medical three-way tube using another syringe to obtain 1 to 2 times white blood cells and 1 to 3 times platelet plasma.
  • a method for extracting platelet plasma from blood including the following steps:
  • the extraction method provided by the comparative example requires the addition of anticoagulant before centrifugation.
  • Two centrifugation operations during centrifugation require more than 20 minutes.
  • the extraction efficiency is low and the speed required for two centrifugation Different, the operation is more cumbersome; before the second centrifugation, it is necessary to extract the uppermost layer and most of the middle layer of plasma.
  • the degree is not easy to grasp during the extraction, and it is easy to mix into the lowermost layer of plasma.

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Abstract

提供了一种自体白细胞和血小板血浆综合提取方法，包括使用常规注射器在冰敷下抽取血液，剪短注射器芯杆并充当离心管进行离心；通过注射器和医用三通管提取血浆，得白细胞和血小板血浆。操作简便，不添加抗凝剂等添加剂。

Description

一种自体白细胞和血小板血浆综合提取方法 技术领域
本发明涉及血浆提取技术领域，具体涉及一种自体白细胞和血小板血浆综合提取方法。
背景技术
富血小板高浓度血浆(PlateletRichPlasma，PRP)，是通过离心血液而得到的含高浓度的血小板血浆。人体的血小板在高浓度状态下，能够产生具有细胞粘合功能的蛋白质(fibrin,fibronectin&vitronectin)，中文名称：纤维连接蛋白、纤粘蛋白，并且当pH值从7.0～7.2降到6.5～6.7时，添加适当的氯化钙，可以促进血小板大量分泌有促进伤口、组织愈合及细胞再生的9种生长因子，所以PRP也叫做“富含生长因子血浆”。PRP具有迅速止血、止痛、加速伤口愈合的作用，可极大程度减轻术后疤痕的形成。另外，由于PRP从患者自身血液中提取，经提取富集处理后，可用于相关疾病的治疗，因而来源丰富，取材方便，可以让身体吸收。自身PRP使用能避免病毒传播和免疫排斥等问题的出现。
但目前在血浆提取过程中，操作步骤繁琐，不利于血浆的快速提取，同时会在提取时添加抗凝剂等添加剂，在血液中添加这些物质，会改变血液的性质，不但使用不便，而且注射后可能会产生过敏、疼痛等问题，影响治疗效果。
发明内容
针对现有技术中的上述不足，本发明提供了一种自体白细胞和血小板血浆综合提取方法，操作简便，可以快速提取得到白细胞和血小板血浆，不添加抗凝剂等添加剂，避免了提取后使用者出现过敏、疼痛等问题，可以直接注射，方便临床使用，并提高了治疗效果。
为实现上述目的，本发明解决其技术问题所采用的技术方案是：提供一种自体白细胞和血小板血浆综合提取方法，包括以下步骤：
(1)使用常规注射器抽取血液，同时将冰袋包裹在常规注射器四周，采血完成后剪短注射器芯杆；
(2)将步骤(1)中剪短后的注射器放入无菌离心套管中，在1～20℃温度下离心2～6min；
(3)步骤(2)中离心后的注射器内血液分为两层，使用另一注射器通过医用三通管提取上层血浆，得白细胞和血小板血浆。
进一步，步骤(1)中剪短注射器芯杆时，在活塞后的凹陷处剪去多余芯杆。
进一步，步骤(2)中离心时，离心力为70～1000g。
进一步，步骤(3)中所得血浆浓度为1～2倍白细胞和1～3倍血小板血浆。
进一步，血液在制备过程中不需要加入抗凝剂等添加剂。
综上所述，本发明具有以下优点：
1、本发明提供的提取方法，在采血时采用冰袋包裹常规注射器，将血液温度立即降低到1～20℃，在此温度下酶的活性降低，血液不易凝固，可以快速提取得到白细胞和血小板血浆；不添加抗凝剂等添加剂，避免了提取后使用者出现过敏、疼痛等问题；可以直接注射，方便临床使用，提高了治疗效果；离心后的上层血浆为含有白细胞和血小板的液态血浆，注射到人体后在注射部位形成凝胶，作用维持7～10天，可以广泛应用于医疗美容、生发、骨性关节炎、运动损伤和牙齿抗衰老等医疗领域。
2、在提取过程中，采血和离心均使用注射器，不需要使用专用的医用耗材，可以降低提取成本，同时在离心后，通过医用三通管提取白细胞和血小板血浆，可以将血浆和红细胞交界面的富白细胞和血小板的成分提取完全，不易混入红 细胞，提取方便快速，节约操作时间，可比目前常用的试管针头提取到更多的白细胞和血小板，并减少红细胞的混入；同时采血针、三通和注射器均为一次性无菌耗材，离心管经过无菌处理，并且密闭，可防止交叉感染，简化了操作步骤。
3、本方案提取的血浆使用时无需激活剂，提取时间相较于常规方法缩短90％，方便直接注射，临床安全性更高，提取物完全来源于自体血浆，其成分和既往的方法提取的成分相同，由于没有添加剂，因此不会出现过敏、排异和感染等风险，更加安全可靠。
具体实施方式
实施例1
一种自体白细胞和血小板血浆综合提取方法，包括以下步骤：
(1)使用常规注射器抽取血液，同时将冰袋包裹在常规注射器四周，采血完成后在活塞后的凹陷处剪去多余芯杆；
(2)将步骤(1)中剪短后的注射器放入无菌离心套管中，在1～20℃温度下离心2min，离心力为70g；
(3)步骤(2)中离心后的注射器内血液分为两层，使用另一注射器通过医用三通管提取上层血浆，得1～2倍白细胞和1～3倍血小板血浆。
实施例2
一种自体白细胞和血小板血浆综合提取方法，包括以下步骤：
(1)使用常规注射器抽取血液，同时将冰袋包裹在常规注射器四周，采血完成后在活塞后的凹陷处剪去多余芯杆；
(2)将步骤(1)中剪短后的注射器放入无菌离心套管中，在1～20℃温度 下离心4min，离心力为500g；
(3)步骤(2)中离心后的注射器内血液分为两层，使用另一注射器通过医用三通管提取上层血浆，得1～2倍白细胞和1～3倍血小板血浆。
实施例3
一种自体白细胞和血小板血浆综合提取方法，包括以下步骤：
(1)使用常规注射器抽取血液，同时将冰袋包裹在常规注射器四周，采血完成后在活塞后的凹陷处剪去多余芯杆；
(2)将步骤(1)中剪短后的注射器放入无菌离心套管中，在1～20℃温度下离心6min，离心力为1000g；
(3)步骤(2)中离心后的注射器内血液分为两层，使用另一注射器通过医用三通管提取上层血浆，得1～2倍白细胞和1～3倍血小板血浆。
对比例
一种从血液中提取血小板血浆的方法，包括以下步骤：
(1)使采集的全血置于含有抗凝剂的容器中，使血液和抗凝剂充分混匀；
(2)使混有抗凝剂的血液置于离心管中，进行第一次离心，离心的转速为2400rpm，时间为4min，使血液基本上分成三层；
(3)将最上层和大部分的中间层抽出并转移到新的离心管中，混合均匀，进行第二次离心，离心的转速为1500rpm，时间为20min；
(4)弃掉离心管上层的血浆，利用剩余的血浆使沉淀的血小板重新悬浮，即得血小板血浆。
对比例所提供的提取方法，相对于本方法而言，在离心前需要加入抗凝剂，在离心时进行两次离心操作，需要20多分钟，提取效率较低，且两次离心所需 转速不同，操作更加繁琐；在第二次离心前，需要抽取最上层和大部分中间层血浆，在抽取时程度不易把握，易混入最下层血浆，所得血小板血浆中混入红细胞，提取效果较差，影响后续的治疗效果。
虽然本发明的具体实施方式对本发明进行了详细地描述，但不应理解为对本专利的保护范围的限定。在权利要求书所描述的范围内，本领域技术人员不经创造性劳动即可作出的各种修改和变形仍属本专利的保护范围。

Claims (5)

  1. 一种自体白细胞和血小板血浆综合提取方法，其特征在于，包括以下步骤：
    (1)使用常规注射器抽取血液，同时将冰袋包裹在常规注射器四周，采血完成后剪短注射器芯杆；
    (2)将步骤(1)中剪短后的注射器放入离心套管中，在1～20℃温度下离心2～6min；
    (3)步骤(2)中离心后的注射器内血液分为两层，使用另一注射器通过医用三通管提取上层血浆，得白细胞和血小板血浆。
  2. 如权利要求1所述的自体白细胞和血小板血浆综合提取方法，其特征在于，步骤(1)中剪短注射器芯杆时，在活塞后的凹陷处剪去多余芯杆。
  3. 如权利要求1所述的自体白细胞和血小板血浆综合提取方法，其特征在于，步骤(2)中离心时，离心力为70～1000g。
  4. 如权利要求1所述的自体白细胞和血小板血浆综合提取方法，其特征在于，步骤(3)中所得血浆为1～2倍白细胞和1～3倍血小板血浆。
  5. 如权利要求1所述的自体白细胞和血小板血浆综合提取方法，其特征在于，血液在制备过程中不需要加入添加剂。
PCT/CN2019/129381 2019-11-21 2019-12-27 一种自体白细胞和血小板血浆综合提取方法 WO2021098011A1 (zh)

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