High-glucose activated mesenchymal stem cell injection and application thereof for diabetic drugs

A technology of diabetes drugs and mesenchymal stem cells, applied in the field of biomedicine, can solve the problems of low efficiency and achieve the effects of improving efficiency, improving insulin resistance, and promoting functional recovery

Inactive Publication Date: 2017-02-01
北京恒峰铭成生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problem of low efficiency of general-purpose mesenchymal stem cell (MSCs) infusion in treating type 2 diabetes (T2D), the present invention provides a method...

Method used

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  • High-glucose activated mesenchymal stem cell injection and application thereof for diabetic drugs
  • High-glucose activated mesenchymal stem cell injection and application thereof for diabetic drugs
  • High-glucose activated mesenchymal stem cell injection and application thereof for diabetic drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Preparation of mesenchymal stem cells

[0031] 1. Preparation of umbilical cord mesenchymal stem cells

[0032]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 suspensi...

Embodiment 2

[0039] Example 2. Preparation of high glucose-activated mesenchymal stem cells

[0040] The mesenchymal stem cells activated by high glucose is to add a certain concentration of glucose solution, after culturing for a certain period of time, measure the number of cultured mesenchymal stem cells, if the concentration of mesenchymal stem cells can reach 5×10 5 ~2×10 6 cells / mL, indicating that the glucose solution has successfully activated mesenchymal stem cells.

[0041] 1. Preparation of umbilical cord mesenchymal stem cells activated by high glucose

[0042] (1)

[0043] When the umbilical cord MSCs prepared in Example 1 were cultured to 40-50% monolayer cells in the fourth generation, glucose solution was added, the final concentration of glucose was 20mmol / L, and continuous culture was carried out for 48 hours; the monolayer cell fusion reached 75%~ At 80%, remove the culture supernatant, digest with 0.25% and 0.1% trypsin-EDTA at 37°C for 1 min, the umbilical cord MSCs...

Embodiment 3

[0054] Example 3. Preparation of High Glucose Activated Mesenchymal Stem Cell Injection

[0055] A high-glucose-activated mesenchymal stem cell injection, which includes the following components: the quantity is 5×10 5 ~2×10 6 Mesenchymal stem cells activated by high glucose 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 solution, the mass volume ratio is 1-5% of 50% glucose injection and the balance is normal saline.

[0056] For example, prepare 100mL of high-glucose-activated 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. 5 mL of normal saline, 85 mL of normal saline, and high glucose-activated MSCs. Except for MSCs activated by high glucose, the rest of the component...

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Abstract

The invention provides a preparation method of high-glucose activated mesenchymal stem cells, and relates to a high-glucose activated mesenchymal stem cell injection and application thereof in preparation of diabetic drugs. The preparation method of the high-glucose activated mesenchymal stem cells comprises the following steps: when culturing an umbilical cord, bone marrow or the mesenchymal stem cells which are extracted separately until fusion reaches 40-50%; adding a glucose solution of which the final concentration is 20-50 mmol/L; culturing for 48+/- 2 hours; and when fusion of the mesenchymal stem cells reaches 75-80%, digesting, neutralizing, washing and collecting the high-glucose activated mesenchymal stem cells. The high-glucose activated mesenchymal stem cell injection comprises the high-glucose activated mesenchymal stem cells, human albumin, compound vitamin, mannitol, glucose and normal saline. The high-glucose activated mesenchymal stem cell injection can be used for treating type-2 diabetes.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation of high-sugar-activated mesenchymal stem cells, a high-sugar-activated mesenchymal stem cell injection and its application in diabetes drugs. Background technique [0002] With the change of modern lifestyle, the incidence of diabetes is getting higher and higher. A recent survey in China found that the incidence of diabetes in China is nearly 10%, that is, the number of people with diabetes is close to 100 million, of which 95% are type 2 Diabetes mellitus (T2D). The pathogenesis of T2D is insulin resistance caused by chronic inflammation caused by obesity, resulting in hyperglycemia and persistent hyperglycemia, which in turn leads to disorders of glucose metabolism and lipid metabolism, thereby causing progressive damage to pancreatic beta cells. Therefore, the treatment of T2D includes two measures: one is to use hypoglycemic drugs and insulin sensitizers...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K9/08A61K47/42A61K47/26A61P3/10C12N5/0775
CPCA61K35/28A61K9/0019A61K9/08A61K47/26A61K47/42C12N5/0663C12N5/0667C12N5/0668C12N2500/34
Inventor 郝好杰李梓源
Owner 北京恒峰铭成生物科技有限公司
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