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Method for preparing platelet lysate and application of platelet lysate

A technology of platelet lysate and platelet, which is applied in the field of cell biology, can solve the problems of great difference in effect, no clear requirements, and small preparation volume, so as to reduce the difference, ensure safety, and increase the content of cytokines.

Inactive Publication Date: 2015-06-03
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Many studies have shown that platelet lysate (PL) for cell culture can be obtained by repeated freezing and thawing, but due to the small amount of preparation, there is no clear requirement for the control of various cytokines in the lysate, resulting in differences in its effect It is very large; there are also reports on the use of machine-collected platelets for large-scale preparation, but no study has been conducted on how to effectively activate platelets to obtain the maximum amount of cytokines. Therefore, how to effectively activate platelets has become a research hotspot

Method used

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  • Method for preparing platelet lysate and application of platelet lysate
  • Method for preparing platelet lysate and application of platelet lysate
  • Method for preparing platelet lysate and application of platelet lysate

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preparation example Construction

[0039] The invention discloses a preparation method of a platelet lysate and its application. Those skilled in the art can learn from the content of this article and appropriately improve the process parameters to realize it. In particular, it should be pointed out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and the relevant personnel can obviously make changes or appropriate changes and combinations to the method and application described herein without departing from the content, spirit and scope of the present invention to realize and Apply the technology of the present invention.

[0040] The preparation method of the platelet lysate provided by the invention and the instrument or reagent used in its application all can be purchased by the market:

[0041] ...

Embodiment 1

[0044] Example 1 Preparation and effect test of platelet lysate

[0045] (1) Platelet sample source, experimental grouping and preparation of platelet lysate

[0046] Select 10 donors of apheresis platelets (from the Chongqing Blood Station of the People's Liberation Army): 7 males and 3 females, aged 18-55 years, all meet the criteria for blood donation (apheresis) without any history of disease. Collect apheresis platelets and collect 10 apheresis plasma (30mL / part) at the same time as the "plasma 0d" group; after apheresis platelets are collected, store them in a shaking box at 22°C for 1 day, and keep 10 apheresis platelets (30mL 10 parts of plasma (30mL / part) were centrifuged to obtain 10 parts of plasma (30mL / part) as the "1d" group, and 10 parts of apheresis platelets (90mL / part) were retained for the preparation of platelet lysate. 10 apheresis platelets (30 mL / part) were centrifuged to obtain 10 plasma (30 mL / part) as the "3d" group, and 10 apheresis platelets (90 mL...

Embodiment 2

[0099] Example 2 Platelet lysate is used for culturing umbilical cord blood mesenchymal stem cells (UMSC)

[0100] According to the ratio of 10%, it was added into the cell culture medium (such as DMEM-F12) to cultivate the umbilical cord blood mesenchymal stem cells (UMSC), and the growth of the cells was observed. Serum-free matrix, calf serum group and platelet lysate (PL) replacement group were set up respectively. A total of 5 groups of cell culture conditions, (1) DMEM / F12 (serum-free, no serum); (2) DMEM / F12+10% FBS; (3) DMEM / F12+10% PL-1 (3d PC-80 ℃ / 37℃ freeze-thaw ultrasonic treatment); (4) DMEM / F12+10%PL-2 (3d PC-ultrasonic treatment); (5) DMEM / F12+10%PL-3 (3d PC-80℃ / freeze-thaw at 37°C). The morphology of UMSC cells cultured for different times under various conditions is shown in Figure 1 to Figure 6 .

[0101] Depend on Figure 1 to Figure 6 The test results show that, compared with fetal bovine serum, the cells cultivated by the platelet lysate prepared by...

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Abstract

The invention relates to the field of cytobiology and in particular relates to a method for preparing platelet lysate and application of the platelet lysate. The preparation method comprises the following steps: oscillating platelets at the temperature of 20-24 DEG C, preserving heat for 1-5 days, treating by adopting a repeated freeze-thaw method and an ultrasonic method, centrifuging, collecting the supernatant, and removing the fibrous proteins, thereby obtaining the platelet lysate. According to the method for preparing the platelet lysate provided by the invention, the content of cell factors in the platelet lysate can be obviously improved, quantitative balance of the cell factors can be realized, the differences among the product batches are reduced, and the effect of stably promoting human-derived cell growth can be achieved.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a preparation method and application of a platelet lysate. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of adult stem cells, which have the commonality of stem cells and strong self-proliferation ability. MSCs have good multi-directional differentiation potential and can be used for the repair of specific tissues and organs; they can participate in the formation of a microenvironment that supports hematopoiesis, promote hematopoietic recovery, and co-transplantation with hematopoietic stem cells can reduce the incidence of GVHD; they have immune regulation and facilitate transplantation The role of MSCs, low immunogenicity, safe and reliable infusion into the human body; therefore, more and more attention has been paid to the clinical application of MSCs. The content of MSCs in bone marrow and umbilical cord blood is very small, about every 10 4 ~10 5 Mono...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N5/071
Inventor 赵树铭王世春范娅涵史春梦
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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