Preparation method of culture medium simulating diabetic foot inflammation micro environment

A technology of diabetic foot and preparation process, which is applied in the direction of culture process, tissue culture, cell culture active agent, etc., can solve the problem of lack of stem cell culture medium, and achieve the effect of good compatibility, appropriate pH and high specificity

Inactive Publication Date: 2018-11-23
广州市番禺区中心医院
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AI Technical Summary

Problems solved by technology

[0006] However, there are not many reports on the influence of diabetic foot inflammatory microenvironment on stem cells, and in vitro cell research on the role of stem cells in diabetic foot inflammatory microenvironment is less, the direct reason is the lack of suitable stem cell culture medium

Method used

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  • Preparation method of culture medium simulating diabetic foot inflammation micro environment

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Embodiment 1

[0017] Use a pipette or a pipette gun to accurately measure 10ml of DMEM / F12 medium, 2ml of fetal bovine serum, 300μl of non-essential amino acids, 300μl of L-glutamine, 200ng of vascular cell adhesion molecule-1, 250ng of tumor necrosis factor α, 200ng of interleukin-6, placed in the same 20ml test tube; vortex the test tube on an automatic vortexer for 10 minutes to fully mix the components; put the fully mixed components into a petri dish and freeze in a 4° refrigerator; According to the number of cells and survival rate, use this medium to adjust the cell inoculation density to 1.0×104 live cells / cm2, add the cell suspension into the culture bottle, shake gently to make the cells evenly distributed; mark it on the culture bottle , transferred to a CO2 incubator with 5% CO2, 37°C, and a saturated humidity of 95%. Observe the state of the cells, and change the medium every other day; when the confluence of the cells reaches 80-90%, they can be subcultured.

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Abstract

The invention relates to the preparation of a stem cell culture medium, in particular to a preparation process of a culture medium simulating a diabetic foot inflammation micro environment. The culture medium consists of the following components: 10ml of DMEM / F12 culture medium, 2ml of fetal calf serum, 300 micro liters of nonessential amino acid, 300 micro liter of L-glutamine, 200ng of vascularcell adhesion molecule-1, 250ng of tumor necrosis factor alpha, and 200ng of interleukin-6. The invention aims at providing a preparation process of the culture medium simulating the diabetic foot inflammation micro environment. The culture medium has specific inflammatory factors causing the diabetic feet such as VCAM1, TNFalpha, IL-6. Mesenchymal stem cells can survive in the culture medium, thecell passage and the cell ultralow-temperature freeze storage can be performed, and the culture medium has important significance for the in-vitro research of stem cells and exosomes in the diabeticfoot inflammation micro environment.

Description

technical field [0001] The invention relates to the preparation of a stem cell culture medium, in particular to a preparation process of a culture medium simulating the inflammatory microenvironment of diabetic foot. Background technique [0002] Sugar In recent years, with the aggravation of population aging and changes in people's diet structure, the incidence of diabetes has risen rapidly. There are more than 415 million diabetic patients in the world, of which China has nearly 100 million diabetic patients. Due to diabetes-induced lower limb ischemia The disease (commonly known as "diabetic foot") can lead to amputation, even life-threatening. It has been reported that 15% of diabetic patients will develop diabetic foot, and the amputation rate of diabetic foot is about 25-50%, which is 40 times that of normal people. The 5-year survival rate is about 50%, so diabetic foot has attracted more and more attention. [0003] Diabetic lower extremity vascular disease is a com...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2500/32C12N2501/2306C12N2501/25C12N2501/58
Inventor 黄晨罗文峰唐郁宽陈汉威庄炜钊李伟奇贺雪平
Owner 广州市番禺区中心医院
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