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Leukocyte extract and preparation method and application thereof

A technology for leukocyte and extract, applied in the field of leukocyte extract and its preparation, can solve the problem of inability to know the effect of beauty and skin care, and achieve the effects of promoting proliferation and metabolism, promoting cleaning, and promoting fusion and proliferation.

Inactive Publication Date: 2015-05-20
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing technology cannot know whether mixing these nucleotides and growth factors can achieve better cosmetic and skin care effects

Method used

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  • Leukocyte extract and preparation method and application thereof
  • Leukocyte extract and preparation method and application thereof
  • Leukocyte extract and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1 Expansion and culture of leukocytes

[0062] Take 80ml of human peripheral blood and mix it with 40ml of normal saline, slowly add it to 60ml of lymphocyte separation medium; centrifuge at 700g, take white blood cells (intermediate buffy coat cells), and inoculate them in culture medium, at 37°C, 5% CO 2 For culturing, the medium was supplemented every 3 to 4 days for 14 days; the medium was RPMI1640 medium, which included 5% FBS and 200 U of interleukin-2. Supplement the medium to the components in the medium and the content of each component is consistent with the initial stage of culture. The experiment was set up in two parallels, and the cell mass was observed regularly, and the results are shown in Table 1:

[0063] Table 1 Changes in cell mass during cell culture

[0064]

[0065] The results showed that after 14 days of culture, 300ml-350ml of culture was obtained, and the number of cells was expanded by 85-100 times.

Embodiment 2

[0066] Example 2 Extraction of Nucleotides in Cultured Leukocytes

[0067] The cultured cultures of Group 1 and Group 2 in Example 1 were taken and centrifuged to obtain cultured leukocytes for the extraction of nucleotides. Another two parts of 80ml human peripheral blood were taken as controls, and white blood cells were separated for nucleotide extraction.

[0068] Add 1 / 5 volume of HClO with a mass fraction of 40% 4 , stir evenly, centrifuge at 1500-2000 rpm, take the supernatant, add KOH to neutralize, stir evenly, take the supernatant by centrifuging at 1500-2000 rpm; add 2- 3 grams of activated carbon, after stirring evenly, extract the activated carbon by suction; add neutral double distilled water to stir, wash, and obtain activated carbon by suction; add alkaline solution, the pH value is 11-13, stir, elute the activated carbon, and obtain by suction filtration The filtrate was neutralized with HCl, the pH was adjusted to neutral, and the isotonicity was adjusted t...

Embodiment 3

[0073] The extraction of growth factor in the embodiment 3 culture fluid

[0074] Take the cultivated cultures of Group 1 and Group 2 in Example 1, separate the leukocytes, and obtain the culture fluid for the extraction of growth factors.

[0075] The culture solution is pre-filtered through a filter membrane with a pore size of 0.45 μm to remove bulky impurities;

[0076] Then filter again through a filter membrane with a pore size of 0.22 μm to remove small volume impurities;

[0077] The obtained filtrate was connected to a Slice200 tangential flow device, and circulated and filtered through a 3kd filter membrane for 3 hours to remove small molecular substances to obtain growth factors.

[0078] After testing, the concentration of the extracted growth factor was 1 mg / mL and 2 mg / mL. The main components of the extract are interleukin 2, interleukin 4, gamma interferon and tumor necrosis factor. Wherein, the mass ratio of interleukin 2, interleukin 4, gamma interferon and...

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Abstract

The invention relates to the technical field of active substances extraction, and particularly relates to a leukocyte extract and a preparation method and application thereof. The leukocyte extract provided by the invention comprises 2.5-7 mu g / mL of nucleotides and 0.3-1.5 mg / mL of proteins. The leukocyte extract can play a role in promoting the fusion and proliferation of epidermis cells, therefore, the leukocyte extract provided by the invention is applied to beauty and skin care products and can achieve whitening, anti-wrinkling and anti-aging effects. The preparation method of the leukocyte extract provided by the invention is implemented through carrying out multiplication culture on leukocytes separated from blood; and compared with the prior art, under the condition of same volume of blood, 85-100 times of nucleotides produced by using the method provided by the invention can be obtained, and a large number of cytokines can be extracted, thereby reducing the demand of blood in traditional methods.

Description

technical field [0001] The invention relates to the technical field of active substance extraction, in particular to a white blood cell extract and its preparation method and application. Background technique [0002] White blood cells (WBC), commonly known as leukocytes, are a very important type of cells in the blood. They have the ability to phagocytize foreign bodies and produce antibodies, heal body damage, and resist pathogen invasion. [0003] The methods for extracting leukocytes as raw materials in the prior art mainly include: firstly, separating leukocytes from blood, and then obtaining leukocyte extracts by lysing and removing impurities. The main components of the product obtained by this method are nucleotides and contain extremely small amounts of cytokines or small molecular polypeptides, and its uses mainly include enhancing immunity, anti-inflammation, antibacterial or anti-pulmonary fibrosis. In the prior art, there is no report about the use of white blo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/98A61K8/64A61Q19/00A61Q19/02A61Q19/08
Inventor 陈海佳王一飞葛啸虎应杰罗二梅
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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