Active mesenchymal stem cell injection for diabetes mellitus infusion

A mesenchymal stem cell and diabetes technology, which is applied in the application of diabetes drugs and the field of active mesenchymal stem cell injection, can solve the problems of low efficiency and achieve the effects of improving efficiency, improving symptoms of kidney and lung complications, and easy quality control

Inactive Publication Date: 2017-08-25
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 low efficiency of general-purpose mesenchymal stem cell (MSCs) infusion in treating type 2 diabetes (T2D), the present invention provides a method for preparing mesenc

Method used

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  • Active mesenchymal stem cell injection for diabetes mellitus infusion
  • Active mesenchymal stem cell injection for diabetes mellitus infusion
  • Active mesenchymal stem cell injection for diabetes mellitus infusion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Preparation of mesenchymal stem cells

[0033] 1. Preparation of umbilical cord mesenchymal stem cells

[0034] 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 suspens...

Embodiment 2

[0041] Example 2. Preparation of mesenchymal stem cells activated by diabetes-like microenvironment

[0042] The mesenchymal stem cells activated by the diabetes-like microenvironment are added with a certain concentration of glucose solution and palmitic acid 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 glucose and palmitic acid solution have successfully activated mesenchymal stem cells. The concentration of palmitic acid is 0.2-0.4mmol / L; the concentration of glucose solution is 20-50mmol / L.

[0043] 1. Preparation of umbilical cord mesenchymal stem cells activated by diabetes-like microenvironment

[0044] (1)

[0045] When the umbilical cord MSC prepared in Example 1 is cultured to 40-50% monolayer cells in the fourth generation, glucose solution and palmitic acid solution are added, the final concentration ...

Embodiment 3

[0052] Example 3. Preparation of Mesenchymal Stem Cell Injection Activated by Diabetic Microenvironment

[0053] An injection of mesenchymal stem cells activated by a DM-like microenvironment, which includes the following components: the quantity is 5×10 5 ~2×10 6 DM-like microenvironment-activated mesenchymal stem cells / 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 The vitamin injection comprises a 50% glucose injection with a mass volume ratio of 1-5% and a balance of normal saline.

[0054]For example, prepare 100mL of DM-like microenvironment-activated mesenchymal stem cell injection, which includes 5mL of human serum albumin stock solution (final concentration of mass-volume ratio is 1%), 2mL of mannitol injection, 3mL of compound vitamin injection, 50% Glucose injection 5mL, saline 85mL, MSC activated by DM-like microenvironment. Except for the MSCs activated by the D...

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Abstract

The invention provides a preparation method of active mesenchymal stem cells, active mesenchymal stem cell injection for diabetes mellitus infusion and application of the injection in preparing drugs for diabetes mellitus. The active mesenchymal stem cells are mesenchymal stem cells activated by diabetes mellitus (DM)-like micro-environment, and the preparation method comprises the steps that when separated and extracted umbilical-cord, bone-marrow or adipose-derived mesenchymal stem cells are cultivated until the fusion rate reaches 40-50%, a glucose solution with the final concentration of 20-50 mmol/L and palmitic acid with the final concentration of 0.2-0.4 mmol/L are added, cultivation is conducted for 46-50 hours, and when the fusion rate of the mesenchymal stem cells reaches 70-80%, the mesenchymal stem cells are digested, neutralized, washed, collected and activated. The active mesenchymal stem cell injection for the diabetes mellitus infusion comprises the mesenchymal stem cells activated by the DM-like micro-environment, human serum albumin, compound vitamins, mannitol, glucose and normal saline. The active mesenchymal stem cell injection for the diabetes mellitus infusion can be used for treating type II diabetes mellitus.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation of mesenchymal stem cells activated by a diabetes-like microenvironment, an active mesenchymal stem cell injection for infusion of diabetes and its application in diabetes drugs. Background technique [0002] Diabetes is a complex metabolic disease. With the change of modern lifestyle, the incidence of diabetes remains high. According to the World Health Organization diabetes data in 2016, 137.6 million adults in China alone suffer from diabetes, with an incidence rate as high as 10%, of which 95% are type 2 diabetes (T2D). The major etiologies of T2D include insulin resistance in target tissues, relative insufficiency of insulin secretion, and subsequent decline in pancreatic β-cell function. Current traditional therapies for T2D include insulin sensitizers to improve hyperglycemia and the use of exogenous insulin to supplement insulin deficiency. β-cell dy...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61K9/08A61K47/42A61P3/10
CPCA61K9/0019A61K9/08A61K35/28A61K47/42C12N5/0663C12N5/0667C12N5/0668C12N2500/34C12N2500/36
Inventor 郝好杰李梓源易军周严恒刘刚陈惠华
Owner 北京恒峰铭成生物科技有限公司
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