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Preparation method and application of conditioned serum rich in cytokines

A cytokine and condition technology, applied in the field of biomedicine, can solve the problems of insignificant efficacy, inability to treat inflammation and degeneration at the same time, low IL-1Ra unit content, etc., to enhance the treatment of tissue inflammation and degeneration, easy operation, and promote healing. Effect

Pending Publication Date: 2022-03-08
CHENGDU QINGKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the content of IL-1Ra units in the conditioned serum obtained by the prior art is low, and the pure conditioned serum rich in anti-inflammatory factors has no obvious efficacy in repairing tissue damage, promoting healing, and restoring functions.
[0006] The platelet-rich plasma (PRP) method is injected into damaged or degenerated tissue parts in an attempt to repair tissue damage and restore function. It has few complications and is a relatively safe treatment method. However, traditionally produced platelets are thick Plasma cannot treat inflammation and degeneration at the same time, therefore, there is room for further improvement

Method used

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  • Preparation method and application of conditioned serum rich in cytokines
  • Preparation method and application of conditioned serum rich in cytokines
  • Preparation method and application of conditioned serum rich in cytokines

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preparation example Construction

[0055] The invention provides a method for preparing IL-1Ra-rich conditioned serum, comprising the steps of:

[0056] Step S1, put fresh blood into glass blood collection tubes containing sterile glass beads, and shake well;

[0057] Step S2, placing the glass blood collection tube filled with whole blood in a constant temperature shaker incubator, incubating with constant temperature shaking, and then centrifuging to remove the red blood cell precipitate, and collecting the supernatant;

[0058] Step S3, filtering the supernatant in step S2 with a sterile filter, and the obtained filtrate is conditioned serum rich in IL-1Ra.

[0059] In step S1, add 30-60 sterile glass beads to the glass blood collection tube per 3 mL of whole blood.

[0060] In step S2, the temperature of the constant temperature shaker incubator is 37° C., the rotation speed is 60-120 r / min, and the shaking incubation time is 12-24 h.

[0061] In step S2, the centrifugation speed is 2500-3500r / min, and th...

Embodiment 1

[0078] Ordinary glass blood collection tube without anticoagulant + shaking culture

[0079] (2) Preparation of materials: prepare ordinary glass blood collection tubes without anticoagulant;

[0080] (2) Preparation of conditional serum rich in IL-1Ra, put fresh blood into ordinary glass blood collection tubes without anticoagulant, shake fully and slowly to ensure that the fresh blood fully contacts the glass blood collection tubes, put the glass blood collection tubes containing blood into Place the blood vessels in a constant temperature shaker incubator at 37°C with a speed of 120r / min, and incubate with constant temperature shaking for 24 hours. Use a centrifuge at a speed of 3500r / min to centrifuge for 10 minutes to remove excess red blood cells. The supernatant serum was filtered through a bacterial filter to obtain IL-1Ra-rich conditioned serum.

Embodiment 2

[0082] Ordinary glass blood collection tube without anticoagulant + shaking culture + glass beads

[0083] (1) Prepare ordinary glass blood collection tubes without anticoagulants and medical grade glass beads, and add 30-60 beads per 3 mL of blood into the glass blood collection tubes;

[0084] (2) Put the fresh blood into the blood collection tube, shake it fully and slowly to ensure that the glass beads are fully in contact with the fresh blood, and place the blood collection tube with the blood and glass beads on a constant temperature shaker at 37°C and a speed of 120r / min In the incubator, carry out constant temperature shaking incubation for 24 hours, and then use a centrifuge at a speed of 3500r / min, centrifuge for 10 minutes to remove excess red blood cell sediment, and finally use a 0.22μm sterile filter to filter the upper serum to obtain IL-1Ra-rich conditioned serum.

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Abstract

The invention discloses a preparation method and application of conditioned serum rich in cytokines, and the preparation method of the conditioned serum comprises the following steps: taking fresh blood, centrifuging, taking PRP part, freezing, thawing and centrifuging to obtain platelet factor-rich plasma, adding sterile glass beads into the rest part, and carrying out constant-temperature oscillation incubation to obtain the conditioned serum rich in the cytokines. And centrifugally filtering to obtain conditioned serum rich in IL-1Ra, and mixing the platelet factor-rich plasma with the conditioned serum rich in IL-1Ra to obtain conditioned serum rich in cytokines. According to the conditioned serum rich in cytokines prepared by the method, platelet factors and IL-1Ra are fully enriched, the content is greatly increased, and meanwhile, the conditioned serum has the effects of repairing tissue damage, recovering functions and treating tissue inflammation and degeneration, and can be used for treating inflammatory arthritis and other inflammatory diseases caused by factors such as trauma, inflammation, infection and degeneration.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a preparation method and application of conditional serum rich in cytokines. Background technique [0002] Arthritis generally refers to an inflammatory disease that occurs in human joints and their surrounding tissues and is caused by inflammation, infection, degeneration, trauma or other factors. The clinical manifestations are redness, swelling, heat, pain, dysfunction and joint deformity of the joints. The patient may lead to joint disability, which greatly affects the quality of life of the patient. [0003] The etiology and pathogenesis of inflammatory arthritis (Inflammatory Arthritis, IA) is not yet fully clear, but a lot of evidence has shown that some cytokines, especially interleukin-1 (IL-1) in the chronic inflammatory process of IA joint pathogenesis In recent years, interleukin-1 receptor antagonist (IL-1Ra) has also been used in the treatment of IA. [0004] ...

Claims

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Application Information

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IPC IPC(8): A61K35/16A61K38/20A61P19/02A61P29/00
CPCA61K35/16A61K38/2006A61P19/02A61P29/00
Inventor 王洁王翔海泉
Owner CHENGDU QINGKE BIOTECH
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