WO2023103744A1 - Procédé d'extraction de facteurs de croissance à partir de plaquettes - Google Patents

Procédé d'extraction de facteurs de croissance à partir de plaquettes Download PDF

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WO2023103744A1
WO2023103744A1 PCT/CN2022/132786 CN2022132786W WO2023103744A1 WO 2023103744 A1 WO2023103744 A1 WO 2023103744A1 CN 2022132786 W CN2022132786 W CN 2022132786W WO 2023103744 A1 WO2023103744 A1 WO 2023103744A1
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platelet
rich plasma
growth
prp
platelets
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PCT/CN2022/132786
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English (en)
Chinese (zh)
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蔡慧思
蔡慧琪
董婷霞
詹华强
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无限发展有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/485—Epidermal growth factor [EGF], i.e. urogastrone
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/49—Platelet-derived growth factor [PDGF]

Definitions

  • the invention relates to the field of biotechnology, in particular to a method for extracting growth factors in platelets.
  • platelet rich plasma that is, PRP is more and more widely used in clinics, including orthopedics, ophthalmology, and plastic surgery.
  • PRP extracted from patients must be fresh and cannot be stored for a long time.
  • the average lifespan of platelets in PRP is usually only 5 to 9 days after extraction, but a normal PRP treatment cycle takes 3 to 6 months.
  • the patient in order to extract PRP, the patient must undergo venipuncture and wait for more than 30 minutes each time receiving PRP treatment.
  • a method for extracting growth factors in platelets disclosed in the embodiments of the present invention can obtain growth group platelet-rich plasma containing a large amount of growth factors.
  • an embodiment of the present invention provides a method for extracting growth factors in platelets, comprising: centrifuging a blood sample with a preset volume to obtain platelet-rich plasma; using a magnetic seat method to remove iron in the platelet-rich plasma ions; the platelet-rich plasma is ultrasonically cracked to obtain the growth group platelet-rich plasma; non-protein inhibitors are added to the growth group platelet-rich plasma; Disinfection and sterilization; and aliquoting and freezing the growth group platelet-rich plasma; wherein, the growth group platelet-rich plasma includes platelet factor 4, ⁇ -thromboglobulin, epidermal growth factor, vascular endothelial growth factor and platelet-derived growth factor .
  • an embodiment of the present invention provides a method for extracting growth factors in platelets, comprising: separating the components of the blood sample to obtain platelet-rich plasma; removing impurities in the platelet-rich plasma; The platelet-rich plasma is subjected to ultrasonic cracking, so that the platelets in the platelet-rich plasma burst and release growth factors to obtain growth group platelet-rich plasma.
  • said growth group platelet-rich plasma includes platelet factor 4, ⁇ -thromboglobulin, epidermal growth factor, vascular endothelial growth factor and platelet-derived growth factor.
  • the extraction method further includes: adding non-protein inhibitors to the platelet-rich plasma of the growth population.
  • the non-protein inhibitor is ethylenediaminetetraacetic acid or ethylene glycol bis(2-aminoethyl ether)tetraacetic acid.
  • the extraction method further includes: performing disinfection and sterilization on the growth group platelet-rich plasma.
  • the sterilizing treatment of the growth group platelet-rich plasma specifically includes: using ultraviolet rays to sterilize the growth group platelet-rich plasma for 25-35 minutes.
  • the extraction method further includes: measuring the protein content in the platelet-rich plasma of the growth group by Bradford method.
  • the extraction method further includes: subpackaging and freezing the growth group platelet-rich plasma.
  • the growth group platelet-rich plasma can be used to prepare skin regeneration products, skin moisturizing products, skin anti-aging products, and products for promoting skin wound healing.
  • the above one or more technical solutions have the following advantages or beneficial effects: obtaining platelet-rich plasma by separating components of the blood sample; removing impurities in the platelet-rich plasma; performing ultrasonic cracking on the platelet-rich plasma,
  • the simple steps of causing platelets in the platelet-rich plasma to burst and release growth factors can obtain growth group platelet-rich plasma with a large amount of growth factors, thereby avoiding multiple blood draws for patients during the PRP treatment cycle, and saving doctors and patients at the same time time and effort, and can reduce the production cost of PRP; in addition, by adding non-protein inhibitors to the platelet-rich plasma and disinfecting and sterilizing the growth group platelet-rich plasma for a preset time, the growth of bacteria can be inhibited, Further extend its shelf life for repeated use by patients; moreover, the obtained growth group platelet-rich plasma is widely used, which can significantly reduce skin pores, increase hemoglobin, skin firmness and elasticity, etc., and can be used to prepare Skin regeneration products, skin moisturizing products, skin
  • Fig. 1 is a schematic diagram of a method for extracting growth factors in platelets provided by an embodiment of the present invention.
  • Fig. 2 is a schematic diagram of a method for extracting growth factors in platelets provided by another embodiment of the present invention.
  • Figure 3 is a schematic diagram of platelet changes measured by a scanning electron microscope in a method for extracting growth factors in platelets provided by an embodiment of the present invention, and the results show that the platelets in the platelet-rich plasma are intact, and the platelets in the growth group platelet-rich plasma are completely broken .
  • Fig. 4 is a bar graph showing the release rate of platelet-released growth factors after platelet-rich plasma is ultrasonically treated for different times in a method for extracting growth factors from platelets according to an embodiment of the present invention.
  • Fig. 5 is a bar graph showing the release rate of each growth factor after adding EDTA and EGTA inhibitors to platelet-rich plasma and performing freeze-thaw cycles in a method for extracting growth factors in platelets provided by an embodiment of the present invention.
  • Fig. 6 is a schematic diagram showing the comparison of the contents of cytokines and growth factors in homologous whole blood WB, platelet-rich plasma and growth group platelet-rich plasma measured by ELISA.
  • Fig. 7 is a schematic diagram showing the comparison of the contents of growth factors EGF, VEGF and PDEGF in platelet-rich plasma and growth group platelet-rich plasma products stored at different temperatures for 1-6 months.
  • Fig. 8 is an experimental diagram and a schematic diagram of the analysis of the cell layer coverage of the wound healing experiment performed on HaCaT cells by platelet-rich plasma and growth group platelet-rich plasma products.
  • Figure 9 is a schematic diagram showing the comparison of transcription levels of COX2, COL1A1, MM2, TNF- ⁇ , COL2A1, COL4A1, COL5A1, COL5A2 and MMP9 genes related to HaCaT cell wound healing by platelet-rich plasma and growth group platelet-rich plasma.
  • Figure 10 is a schematic diagram of the comparison of the changes in the pores of the cheeks before and after 6 months of application of platelet-rich plasma or growth group platelet-rich plasma to the skin of volunteers.
  • Figure 11 shows the facial skin moisture, water retention capacity, whitening effect, hemoglobin level, oil level, skin gloss, firmness and elasticity of volunteers who received platelet-rich plasma or growth group platelet-rich plasma for 6 months. Schematic diagram of the comprehensive evaluation results of the skin before and without use.
  • FIG. 12 is a schematic diagram of experiments comparing the T0, T4, and T8 cell layer coverages of HaCaT cells in growth group platelet-rich plasma and platelet-rich plasma of eel blood.
  • Fig. 13 is a schematic diagram of experiments comparing T0, T10, and T24 cell layer coverages of HaCaT cells in the growth group platelet-rich plasma and platelet-rich plasma of deer blood in the wound healing experiment.
  • the “growth group” mentioned in the embodiment of the invention refers to various growth factors contained in platelets in the blood, which can be collected by ultrasonically destroying the platelet membrane and then fully released, mainly including platelet-derived growth factor (PDGF), Vascular endothelial cytokine (VEGF) and epidermal growth factor (EGF), etc.
  • PDGF platelet-derived growth factor
  • VEGF Vascular endothelial cytokine
  • EGF epidermal growth factor
  • the blood (or blood sample) used is from human or other animals.
  • the prepared growth population platelet-rich plasma can be used for the individual from whom the blood was drawn.
  • Embodiment 1 of the present invention proposes a method for extracting platelet growth factor in blood.
  • the method for extracting growth factors in platelets includes, for example, the following steps S11 to S21.
  • the growth group platelet-rich plasma includes platelet factor 4 (PF4), ⁇ -thromboglobulin ( ⁇ -TG), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) .
  • PF4 platelet factor 4
  • ⁇ -TG ⁇ -thromboglobulin
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • PDGF platelet-derived growth factor
  • the preset volume of blood sample mentioned in step S11 is, for example, a blood sample of 30ml-60ml.
  • the referenced blood can be of human or other animal origin.
  • the centrifugation mentioned can be understood as the separation of components with different specific gravity in blood by means of centrifugal force.
  • the ultrasonic lysis mentioned in S15 can be understood as the complete rupture of platelets to release the internal growth factors and cytokines, as shown in Figure 3 .
  • the preset time mentioned in step S19 is, for example, 30 minutes.
  • a blood sample with a preset volume is centrifuged to obtain platelet-rich plasma, and the platelet-rich plasma is removed by using a magnetic seat method.
  • Iron ions of the platelet-rich plasma were ultrasonically cracked to obtain growth group platelet-rich plasma, non-protein inhibitors were added to the growth group platelet-rich plasma, and ultraviolet rays were used to preset the growth group platelet-rich plasma
  • Timely disinfection and sterilization, as well as aliquoting and freezing the growth group platelet-rich plasma can obtain growth group platelet-rich plasma with a large amount of growth factors and a longer shelf life.
  • the method for extracting the growth factor in platelets further includes, for example: measuring the protein content in the platelet-rich plasma of the growth group by using the Bradford method.
  • Embodiment 1 of the present invention proposes a method for extracting growth factors in platelets.
  • the method for extracting growth factors in platelets includes, for example, the following steps S31 to S35.
  • the blood sample mentioned in step S31 may be from human beings or from other animals.
  • the separation mentioned is, for example, to separate blood components by centrifugation, but it is not limited here, as long as the same or similar functions can be achieved.
  • the impurities mentioned in step S33 are, for example, iron ions, which are removed by, for example, a magnetic seat method.
  • the ultrasonic lysis mentioned in step S35 can be understood as complete rupture of platelets to release internal growth factors and cytokines, as shown in FIG. 3 .
  • the growth group platelet-rich plasma mentioned in step S35 includes, for example, platelet factor 4 (PF4), ⁇ -thromboglobulin ( ⁇ -TG), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF) and platelet Derived growth factor (PDGF).
  • PF4 platelet factor 4
  • ⁇ -TG ⁇ -thromboglobulin
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • PDGF platelet Derived growth factor
  • the time of ultrasonic lysis will affect the release rate of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF).
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • PDGF platelet-derived growth factor
  • the method for extracting growth factors in platelets further includes, for example: adding non-protein inhibitors to the platelet-rich plasma of the growth group.
  • non-protein inhibitors are, for example, ethylenediaminetetraacetic acid (EDTA) or ethylene glycol bis (2-aminoethyl ether) tetraacetic acid (EGTA), which are inhibitors that do not cause skin allergies, and specific inhibitors
  • EDTA ethylenediaminetetraacetic acid
  • EGTA ethylene glycol bis (2-aminoethyl ether) tetraacetic acid
  • specific inhibitors are not limited to EGTA and EDTA, as long as the same effect can be achieved, the specific concentration is not particularly limited, and can be any effective concentration that can inhibit microorganisms.
  • the effect of adding non-protein inhibitors here is not only to prevent skin irritation, but also to inhibit microbial growth to further extend its shelf life.
  • the addition of inhibitors EDTA and EGTA can promote the release of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) in platelets.
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • PDGF platelet-derived growth factor
  • the freeze-thaw cycle refers to freezing and then thawing, that is, freezing once and thawing once, which is different from the "repeated freezing and thawing" of the traditional technology (ie, freezing and thawing repeatedly).
  • the method for extracting growth factors in platelets further includes, for example: disinfecting and sterilizing the growth group platelet-rich plasma.
  • the disinfection and sterilization treatment of the growth group platelet-rich plasma is specifically: using ultraviolet rays to disinfect and sterilize the growth group platelet-rich plasma for 25-35 minutes, preferably 30 minutes, so as to achieve sterilization, and then prolong its life.
  • the role of shelf life It is worth noting that after the platelet-rich plasma of the growth group is sterilized, it needs to be immediately divided into sterilized microcentrifuge tubes, freeze-dried, and stored in a -80°C low-temperature refrigerator for later use.
  • the method for extracting the growth factor in platelets further includes, for example: measuring the protein content in the platelet-rich plasma of the growth group by using the Bradford method.
  • the protein content of the test solution is measured by the Bradford method, so that the samples distributed in each sterilized microcentrifuge tube have about the same amount of protein, such as 1 mg of protein. Note here that before using the growth group platelet-rich plasma sample, it is necessary to add sterile saline (0.9% NaCl) to dissolve the protein.
  • the method for extracting growth factors in platelets further includes, for example: subpackaging and freezing the growth group platelet-rich plasma, so as to prolong its shelf life and use it multiple times.
  • the growth group platelet-rich plasma obtained based on the above method can be used to prepare skin regeneration products, skin moisturizing products, skin anti-aging products, and products for promoting skin wound healing.
  • each component of the blood sample is separated to obtain platelet-rich plasma, and impurities in the platelet-rich plasma are removed;
  • the simple steps of performing ultrasonic cracking of the platelet-rich plasma to make the platelets in the platelet-rich plasma burst and release growth factors can obtain growth group platelet-rich plasma with a large amount of growth factors, thereby avoiding multiple times of PRP treatment cycles for patients.
  • Blood drawing saves the time and energy of doctors and patients, and can reduce the production cost of PRP; in addition, by adding non-protein inhibitors to the growth group platelet-rich plasma, the steps of disinfection and sterilization of the growth group platelet-rich plasma , can inhibit the growth of bacteria, and greatly prolong the shelf life of the growth group platelet-rich plasma; in addition, the growth group platelet-rich plasma can be subpackaged and frozen for users to take multiple times; moreover, based on the extraction method of the growth factor in the platelets, the The growth group platelet-rich plasma is widely used, and it can be applied to the preparation of skin regeneration products, skin moisturizing products, skin anti-aging products, and skin wound healing products.
  • cytokines and growth factors include: platelet factor-4 (PF 4), ⁇ -thromboglobulin ( ⁇ -TG), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) content, its specific detection steps are as follows:
  • PRP Plus contains significantly more cytokines and growth factors.
  • the amount of ⁇ -TG, EGF and VEGF released in PRP Plus is almost twice that of whole blood WB or PRP, which is significantly better than PRP and WB.
  • This result shows that the present invention can obtain more cytokines and growth factors, such as growth factors such as PF4, ⁇ -TG, EGF, VEGF, PDGF, by preparing PRP Plus.
  • These active growth factors can be quickly detected by ELISA, so they can be used as technical quality control parameters of PRP Plus.
  • the PRP and PRP Plus prepared by the above-mentioned embodiment one or embodiment two were stored at different temperatures for 6 months by detecting each growth factor: epidermal growth factor (EGF), vascular endothelial growth factor (VEGF) and platelet-derived Growth factor (PDGF) content to determine the stability of PRP Plus.
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • PDGF platelet-derived Growth factor
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • PDGF platelet-derived growth factor
  • the PRP and PRP Plus obtained in the above-mentioned embodiment one or embodiment two are applied to the wound healing experiment, and the specific steps are as follows:
  • FIG. 8 (left) for a schematic diagram of the cell layer coverage of HaCaT cells, which correspond to the blank control, PRP, positive control (VEGF) and PRP Plus from left to right, and T0 and T8 from top to bottom. After the cells were cultured for 8 hours, the morphological observation showed that the wound healing rate of cells treated with PRP Plus was significantly higher than that of PRP, at least doubled.
  • Fig. 8 (right) is an analysis diagram of the cell layer coverage of HaCaT cells, which is the coverage ratio of each group of experiments compared with the blank control after 8 hours. It can be concluded that PRP Plus technology can speed up wound healing.
  • HaCaT cells were planted in a 100 mm culture dish, and blank control, positive control (VEGF), 10% PRP, 10% PRP Plus were added and cultured for 24 hours.
  • VEGF positive control
  • 10% PRP 10% PRP Plus
  • SYBR Green performs real-time fluorescence quantitative polymerase chain reaction on equal amounts of each cDNA sample to detect the relative transcription levels of COX2, COL1A1, MM2, TNF- ⁇ , COL2A1, COL4A1, COL5A1, COL5A2 and MMP9 genes, using GAPDH gene level as an internal reference.
  • the PRP and PRP Plus prepared in the above-mentioned embodiment 1 or embodiment 2 are applied to the individual facial skin.
  • 20 healthy volunteers were used to conduct a 6-month experiment on autologous facial skin, using instruments to detect the skin before and after using PRP and PRP Plus products.
  • the age distribution range of the 20 healthy volunteers was: : 20 to 75 years old; 7 males and 13 females. They were randomly divided into two groups, with 10 subjects in each of PRP and PRP Plus.
  • the test period is six months, using PRP or PRP Plus once a month.
  • the room temperature is 22 ⁇ 2°C
  • the relative humidity is controlled at 50 ⁇ 5%
  • the test sites forehead, left cheek, right cheek, and chin.
  • VISIA Canfield Scientific Inc, NJ
  • the parameters of the skin surface are tested, eg with the Courage+Khazaka Electronics GmbH (C+K) system (Cologne, Germany).
  • C+K Courage+Khazaka Electronics GmbH
  • CK skin tester moisture test probe, water loss test probe, melanin test probe, oil test probe, gloss test probe, elasticity test probe to test the skin hydration, water retention, whitening effect, Erythema level, sebum level, gloss and elasticity were tested.
  • a series of skin parameters were evaluated on 20 subjects using 6 probes connected to the C+K skin testing system, including skin moisture, water retention capacity (testing water loss rate), whitening effect (reduction of melanin content), erythema Level (level of hemoglobin content), skin oil level, radiance, firmness and elasticity of the skin.
  • PRP Plus has a stronger effect of shrinking pores, and can improve skin moisturizing ability, reduce sebum, and increase skin rosiness, which can be applied to autologous skin wound healing, skin regeneration, skin whitening, Or skin antioxidant/anti-aging. It can also further prepare articles related to skin regeneration, skin moisturizing, skin anti-aging or assisting skin wound healing according to its efficacy.
  • eel blood was used to prepare PRP and PRP Plus based on the preparation method provided in Example 1 or Example 2 above, so as to perform wound healing experiments on HaCaT cells. Comparing the cell layer coverage of PRP and PRP Plus in eel blood on HaCaT cells in wound healing experiments at T0, T4, and T8, it can be seen from the figure that PRP Plus in eel blood is significantly better than PRP in eel blood in promoting wound healing.
  • deer blood was used to prepare PRP and PRP Plus, so as to conduct wound healing experiments on HaCaT cells. Comparing the cell layer coverage rate of PRP and PRP Plus of deer blood on HaCaT cells in wound healing experiments at T0, T10, and T24, it can be seen from the figure that PRP Plus of deer blood is significantly better than PRP of deer blood in promoting wound healing.

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Abstract

L'invention concerne un procédé d'extraction de facteurs de croissance à partir de plaquettes, comprenant les étapes consistant à : centrifuger un échantillon de sang d'un volume prédéfini pour obtenir un plasma riche en plaquettes ; éliminer les ions fer dans le plasma riche en plaquettes au moyen d'un procédé à base magnétique ; réaliser une lyse ultrasonore sur le plasma riche en plaquettes pour obtenir un plasma riche en plaquettes de groupe de croissance ; ajouter des inhibiteurs non protéiques dans le plasma riche en plaquettes de groupe de croissance ; utiliser une lumière ultraviolette pour désinfecter et stériliser le plasma riche en plaquettes de groupe de croissance pendant une période de temps prédéfinie ; aliquoter et congeler le plasma riche en plaquettes de groupe de croissance. Le plasma riche en plaquettes de groupe de croissance comprend le facteur plaquettaire 4, la β-thromboglobuline, le facteur de croissance épidermique, le facteur de croissance endothéliale vasculaire et le facteur de croissance dérivé des plaquettes. Le plasma riche en plaquettes de groupe de croissance ayant une grande quantité de facteurs de croissance peut être obtenu au moyen d'étapes simples, et peut être utilisé pour préparer des produits de régénération de la peau, des produits d'hydratation de la peau, des produits anti-âge de la peau et des produits pour favoriser la cicatrisation des plaies de la peau.
PCT/CN2022/132786 2021-12-07 2022-11-18 Procédé d'extraction de facteurs de croissance à partir de plaquettes WO2023103744A1 (fr)

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CN112341534A (zh) * 2020-07-29 2021-02-09 璟骐生物科技(深圳)有限公司 血小板生长因子及其提取方法和应用

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005105121A1 (fr) * 2004-05-05 2005-11-10 Synthes Gmbh Utilisation de plaquettes ou de plasma riche en plaquettes (prp)
US20170157018A1 (en) * 2013-09-17 2017-06-08 Bestop Group Holdings Limited Growth factor concentrate and the use thereof
CN106995795A (zh) * 2016-01-26 2017-08-01 李建业 基于富血小板血浆的细胞处理方法、试剂盒及冻干粉
CN110891582A (zh) * 2017-06-16 2020-03-17 里尔大学中心医院 制备混合人血小板裂解液的方法、混合人血小板裂解液及其用于治疗神经系统疾病的用途
CN107446036A (zh) * 2017-09-25 2017-12-08 浙江中医药大学 一种血小板细胞因子浓集物的富集方法
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