A preparation method of microspheres loaded with platelet lysate

A platelet lysate, platelet technology, applied in the direction of blood/immune system cells, fixed on/in organic carriers, bone/connective tissue cells, etc. question

Active Publication Date: 2021-09-14
天晴干细胞股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to provide a method for preparing microspheres loaded with platelet lysate in order to solve the problem of large consumption of platelet lysate or poor adsorption effect of microcarriers in the existing cell culture

Method used

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  • A preparation method of microspheres loaded with platelet lysate
  • A preparation method of microspheres loaded with platelet lysate
  • A preparation method of microspheres loaded with platelet lysate

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Experimental program
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specific Embodiment approach 1

[0024] Specific embodiment one: In this embodiment, a method for preparing microspheres loaded with platelet lysate is carried out according to the following steps:

[0025] 1. Add basement membrane proteoglycan solution to the platelet lysate rich in CD41+ and CD81+ microcapsules, and stir evenly while adding to obtain composition A; wherein the platelet lysate and basement membrane protein rich in CD41+ and CD81+ microcapsules The volume ratio of polysaccharide solution is (3.7-5.5):(0.8-2.4);

[0026] 2. At room temperature, add the sodium alginate solution to composition A, and mix well to form composition B; wherein the volume ratio of the sodium alginate solution to the platelet lysate rich in CD41+ and CD81+ microcapsules is (3.0- 5.0):(3.7-5.5);

[0027] 3. Add composition B to a spray gun equipped with a porous nozzle, use nitrogen as the power, and spray into 3-10% CaCl at 4°C with 90rpm magnetic stirring at a speed of 1-30m / s 2 In aqueous solution, microspheres wi...

specific Embodiment approach 2

[0032] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the preparation method of the platelet lysate rich in CD41+, CD81+ microcapsules in step one is:

[0033] 1. Use a focused extracorporeal shock wave instrument with a frequency of 15Hz and an energy-flux density of 0.15-0.20mJ / mm 2 , after shocking platelet-rich plasma or platelet-rich saline 200-500 times, at 37°C CO 2 Incubate in the incubator for 10-15 minutes; then at a frequency of 15Hz, the energy flux density is 0.20-0.25mJ / mm 2 , shock 700-1000 times, at 37°C CO 2 Incubate in the incubator for 20-30 minutes; then at a frequency of 15Hz, the energy flux density is 0.25-0.30mJ / mm 2 , shock 1500-2000 times, at 37°C CO 2 Incubate in an incubator for 10 minutes to obtain a suspension;

[0034] 2. Put the suspension in step 1 into a sealed freezing tube, put it into liquid nitrogen to freeze for 20-40 minutes, take it out and put it in a water bath to shake and melt, re...

specific Embodiment approach 3

[0036] Specific embodiment 3: The difference between this embodiment and specific embodiment 1 or 2 is that the platelet concentration in platelet-rich plasma or platelet-rich saline is (6-15)×10 5 pieces / μl. It is the same as the specific embodiment 1 or 2.

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Abstract

A method for preparing microspheres loaded with platelet lysate. The present invention relates to a method for preparing microspheres loaded with platelet lysate. The purpose of the invention is to solve the problem of large consumption of platelet lysate or poor adsorption effect of microcarriers in existing cell culture problem, the present invention includes preparation of platelet lysate, preparation of microsphere components, preparation of microspheres loaded with platelet factors and microcapsules. The microspheres prepared by the present invention have a very strong ability to carry osteoblasts. In the case of osteoblasts: microspheres = 500:1, most of the cells adhere to the microspheres, and only a few cells are scattered in the culture medium. The mesenchymal stem cells grow well on the microspheres, the cell viability exceeds 95%, the doubling time is short, and the positive expression rate in terms of purity exceeds 95%. The invention is applied in the field of biotechnology.

Description

technical field [0001] The invention relates to a method for preparing microspheres loaded with platelet lysate. Background technique [0002] Platelets have been regarded as non-functional cell fragments in the blood for a long time. It was not until 1882 that the Italian physician Bizzozero discovered that they played an important role in the hemostasis process after vascular injury, and it was first proposed that the name of platelets began. Explore the process of platelet coagulation, hemostasis, and maintenance of capillary wall integrity. It is now recognized that platelets have important functions. [0003] Platelets are biologically active small pieces of cytoplasm that are lysed and shed from the cytoplasm of mature megakaryocytes in the bone marrow. They are small in size, 2-3 microns in diameter, and have no nucleus. Platelets have a specific morphological structure and biochemical composition, and have a relatively constant number in normal blood (for example, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775C12N5/078C12N11/10
CPCC12N5/0644C12N5/0665C12N11/10
Inventor 张怡吕秀明牛春玮刘艳青
Owner 天晴干细胞股份有限公司
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