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Preparation method and application of a kind of anti-chronic inflammation mesenchymal stem cells

A technology of mesenchymal stem cells and chronic inflammation, applied in the field of anti-chronic inflammation application and mesenchymal stem cell injection for anti-chronic inflammation, can solve the problems of low efficiency and uncertain treatment effect, achieve optimal treatment plan, enhance anti-inflammatory and anti-inflammatory effects. The effect of inflammation, the effect of easy quality control

Active Publication Date: 2020-06-23
恒峰铭成生物科技(上海)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Aiming at the problems that MSCs are affected by the internal environment, the therapeutic effect is uncertain and the efficiency is not high, the present invention provides a preparation method of mesenchymal stem cells against chronic inflammation, an injection of mesenchymal stem cells against chronic inflammation and its application for anti-chronic inflammation

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  • Preparation method and application of a kind of anti-chronic inflammation mesenchymal stem cells
  • Preparation method and application of a kind of anti-chronic inflammation mesenchymal stem cells

Examples

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Effect test

Embodiment 1

[0027] Example 1. Preparation of umbilical cord mesenchymal stem cells

[0028]Operate in an ultra-clean workbench, take fresh full-term healthy fetal umbilical cord, wash it with normal saline containing 2 times 100U / mL penicillin and streptomycin to remove blood stains, peel off Walton’s jelly, and cut it into pieces less than 1 cubic millimeter Shape, spread evenly, slowly add 10mL serum-free medium, and culture in a 37°C, 5% carbon dioxide, saturated humidity incubator. After the primary cells were cultured for 7 days, the serum-free medium was replaced and cultured until the cells reached 70% confluence, the serum-free medium was removed, and trypsin-sodium edetate ( EDTA) for 1 min, the umbilical cord MSCs shrank and detached from the bottle wall. At this time, the previously removed culture supernatant was added to neutralize the trypsin-EDTA solution, and gently blown with a pipette, and the umbilical cord MSC suspension was transferred to a 50 mL centrifuge. Tube. C...

Embodiment 2

[0031] Example 2. LPS hypoxic pretreatment of umbilical cord mesenchymal stem cells

[0032] The anti-chronic inflammation mesenchymal stem cells are LPS hypoxic pretreated mesenchymal stem cells, and the preparation method of the anti-chronic inflammation mesenchymal stem cells includes the following steps: a) culturing the mesenchymal stem cells to 70-80 % fusion, remove the culture medium in the mesenchymal stem cell culture dish, wash the mesenchymal stem cells with phosphate buffer; b) add serum-free medium containing 50-200ng / mL lipopolysaccharide (LPS); then c) Adjusting the hypoxic state to an oxygen content of 5-10%, and culturing the mesenchymal stem cells in a hypoxic state for 16-18 hours, measuring the number of cultured mesenchymal stem cells, if the number of mesenchymal stem cells can reach 5 ×10 5 ~2×10 6 cells / mL, indicating effective LPS and hypoxia pretreated mesenchymal stem cells.

[0033] (1)

[0034] Inoculate 1.5×10 per 15 cm cell culture dish 6 U...

Embodiment 3

[0039] Example 3. Preparation of Mesenchymal Stem Cell Injection against Chronic Inflammation

[0040] An anti-chronic inflammation mesenchymal stem cell injection, which includes the following components: the quantity is 5×10 5 ~2×10 6 LPS hypoxic pretreated mesenchymal stem cells per mL, the mass volume ratio is 1-5% human serum albumin, the mass volume ratio is 1-8% mannitol injection, and the mass volume ratio is 1-5% compound Vitamin injection, 50% glucose injection with a mass volume ratio of 1-5% and normal saline as the balance, wherein the mesenchymal stem cells against chronic inflammation are lipopolysaccharide and hypoxic pretreated mesenchymal stem cells.

[0041] For example, prepare 100mL of anti-chronic inflammation mesenchymal stem cell injection, which includes 5mL of human serum albumin stock solution (the final concentration of mass volume ratio is 1%), 2mL of mannitol injection, 3mL of compound vitamin injection, and 50% glucose injection. MSCs pretreate...

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Abstract

The invention provides a preparing method and application of anti-chronic inflammation mesenchymal stem cells. The preparing method comprises the steps of culturing mesenchymal stem cells obtained through separation and extraction till the fusion rate reaches 70-80%, removing a culture medium in a mesenchymal stem cell culture dish, and flushing the mesenchymal stem cells with phosphate buffer; then adding a serum-free medium containing 50-200 ng / mL lipopolysaccharide; and adjusting the hypoxia state till oxygen content becomes 5-10%, and culturing the mesenchymal stem cells for 16-18 h in the hypoxia state. The anti-chronic inflammation mesenchymal stem cells can be used for treating chronic inflammation.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation method of anti-chronic inflammation mesenchymal stem cells, an anti-chronic inflammation mesenchymal stem cell injection and its application for anti-chronic inflammation. Background technique [0002] Chronic inflammation can cause a variety of diseases, such as diabetes, cardiovascular disease, etc., and is the common pathogenesis of many diseases. The pathogenesis of type 2 diabetes is insulin resistance caused by chronic inflammation caused by obesity, resulting in hyperglycemia, and persistent hyperglycemia, which in turn leads to disorders of glucose and lipid metabolism, resulting in progressive damage to pancreatic β cells. Obesity causes cells to swell, and the body's immune system recognizes them as abnormal cells, modifies them, and causes excess inflammation in the body, leading to diabetes. In cardiovascular diseases, vascular endothelial cells a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61K35/28A61K9/08A61K47/42A61K47/26A61P29/00
CPCA61K9/0019A61K9/08A61K35/28A61K47/26A61K47/42C12N5/0665C12N2500/34C12N2500/90
Inventor 郝好杰李梓源易军周严恒陈惠华
Owner 恒峰铭成生物科技(上海)有限公司
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