Human skin flbroblast culture fluid and cultural method

A fibroblast and culture method technology, applied in the field of human skin fibroblast culture medium and culture, can solve problems such as affecting cell growth rate, insufficient oxygen supply, mechanical damage, etc., and achieves inhibition of human skin fibroblast apoptosis, Effect of improving density and functional status, prolonging in vitro survival time

Inactive Publication Date: 2017-05-10
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 2. Relatively insufficient oxygen supply
This lack of oxygen can not only directly cause cell damage and affect the growth rate of cells, but also promote the anaerobic glycolysis of glucose in the medium, increase the production of lactic acid, make the pH value in the medium drop sharply, and affect the human body. Skin fibroblasts cause further damage
[0006] 3. Mechanical damage
However, under the action of many forces (such as the shear force generated by turbulence and air bubbles, and the collision between microcarriers) during cell culture, the cells attached to the microcarriers will be damaged, inhibit cell growth, and cause cell death

Method used

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  • Human skin flbroblast culture fluid and cultural method
  • Human skin flbroblast culture fluid and cultural method
  • Human skin flbroblast culture fluid and cultural method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Large-scale cultivation of human skin fibroblasts using the culture medium disclosed in the present invention

[0055] 1. Preparation of culture medium

[0056] Test group: in 1L DMEM (high sugar type) medium containing 60mL fetal bovine serum, 35mmol HEPES, 60mg total flavonoid extract of rose fruit, 20mg astragalus polysaccharide and 10 -7 mmol insulin.

[0057] Control group 1: 60 mL of fetal bovine serum and 35 mmol of HEPES were contained in 1 L of DMEM (high glucose type) medium.

[0058] Control group 2: 60 mL of fetal bovine serum, 35 mmol of HEPES, and 120 mg of Rosa rosea total flavonoids extract were contained in 1L of DMEM (high glucose type) medium.

[0059] Control group 3: 60 mL of fetal bovine serum, 35 mmol of HEPES, and 40 mg of astragalus polysaccharide were contained in 1 L of DMEM (high glucose type) medium.

[0060] Control group 4: In 1L DMEM (high glucose type) medium containing 60mL fetal bovine serum, 35mmol HEPES, 2×10 - 7 mmol...

Embodiment 2

[0067] Example 2: Large-scale cultivation of human skin fibroblasts using the culture medium disclosed in the present invention

[0068] Test group: in 1L DMEM (high-glucose type) medium containing 60mL fetal bovine serum, 35mmol HEPES, 50mg total flavonoid extract of rose fruit, 30mg astragalus polysaccharide and 10 -8 mmol insulin.

[0069] Control group 1: 60 mL of fetal bovine serum and 35 mmol of HEPES were contained in 1 L of DMEM (high glucose type) medium.

[0070] Control group 2: 60 mL of fetal bovine serum, 35 mmol of HEPES, and 100 mg of Rosa rosea total flavonoids extract were contained in 1 L of DMEM (high glucose type) medium.

[0071] Control group 3: 60 mL of fetal bovine serum, 35 mmol of HEPES, and 60 mg of astragalus polysaccharide were contained in 1 L of DMEM (high glucose type) medium.

[0072] Control group 4: In 1L DMEM (high glucose type) medium containing 60mL fetal bovine serum, 35mmol HEPES, 2×10 - 8 mmol insulin.

[0073] According to the sam...

Embodiment 3

[0075] Example 3: Large-scale cultivation of human skin fibroblasts using the culture medium disclosed in the present invention

[0076] Test group: in 1L DMEM (high sugar type) medium containing 60mL fetal bovine serum, 35mmol HEPES, 80mg thorn rose fruit total flavonoid extract, 40mg astragalus polysaccharide and 10 -6 mmol insulin.

[0077] Control group 1: 60 mL of fetal bovine serum and 35 mmol of HEPES were contained in 1 L of DMEM (high glucose type) medium.

[0078] Control group 2: 60 mL of fetal bovine serum, 35 mmol of HEPES, and 160 mg of Rosa rosea total flavonoids extract were contained in 1L of DMEM (high glucose type) medium.

[0079] Control group 3: 60 mL of fetal bovine serum, 35 mmol of HEPES, and 80 mg of astragalus polysaccharide were contained in 1 L of DMEM (high glucose type) medium.

[0080] Control group 4: In 1L DMEM (high glucose type) medium containing 60mL fetal bovine serum, 35mmol HEPES, 2×10 - 6 mmol insulin.

[0081] According to the sam...

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Abstract

The invention relates to the field of biotechnology, and specifically relates to a human skin flbroblast culture fluid. The human skin flbroblast culture fluid disclosed by the invention is prepared from a culture medium, fetal calf serum, HEPES, a rose hip flavonoid extract, astragalus polysaccharide and insulin. The culture fluid has a good effect for large-scale culture of human skin flbroblast.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a human skin fibroblast culture solution and a culture method. Background technique [0002] With the maturity of human skin fibroblast isolation and culture technology and the development of biomedical engineering and other technologies, bioartificial skin based on cultured human skin fibroblasts has gradually developed. People have always hoped to adopt this method of skin support in vitro To replace the damaged skin function of the patient, and create conditions for the patient to wait for skin transplantation or self-skin regeneration. At present, it is believed that at least 10 10 Cells of the above order of magnitude, and under the condition of ensuring cell activity, the more the number of human skin fibroblasts, the better the support and treatment effect. Therefore, the large-scale culture technology of human skin fibroblasts in vitro has become a bioartificial skin technol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N2500/76C12N2501/33C12N2501/90
Inventor 陈海佳葛啸虎王一飞刘帅王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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