Preparing method of immune-cell-derived biological-beautifying raw materials and products thereof

A technology of immune cells and raw materials, applied in the direction of cosmetic preparations, dressing preparations, skin care preparations, etc., which can solve problems such as functional and biological activity limitations, skin allergies, and ineffectiveness

Active Publication Date: 2016-10-12
ZICHENG RUISHENGHUI BEIJING BIOTECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since these cytokines are expressed in lower cells such as Escherichia coli or yeast, they lack the post-translational modification of protein transcription in mammalian cells, including processes such as glycosylation, methylation and terminal modification, and their functions and Biological activity is very limited
Therefore, when the cytokines prepared by fermentation in E. coli or yeast are used in cosmetics, many of them may not have high biological activity or even be ineffective, or cause skin allergies due to immunogenicity problems.

Method used

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  • Preparing method of immune-cell-derived biological-beautifying raw materials and products thereof
  • Preparing method of immune-cell-derived biological-beautifying raw materials and products thereof
  • Preparing method of immune-cell-derived biological-beautifying raw materials and products thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Culture of human immune cells

[0055] Collect 100ml of peripheral blood from volunteers (with whom they signed an informed consent form), dilute it with 1×PBS (pH value 7.4), and obtain mononuclear cells after density gradient centrifugation of lymphocyte separation medium, using 1000IU / ml cytokines Interferon γ, 50ng / ml anti-human CD3 monoclonal antibody, 100IU / ml interleukin 1α and 500IU / ml interleukin 2 (all purchased from Germany Miltenyi company) in serum-free medium AIM-V (containing 50μg / ml hydrocortisone pine, 10 μg / ml ascorbic acid and 50 μg / ml human insulin, all purchased from American Life Company) and continuously cultured for 20 days to obtain 5.0×10 9 Cytokines induce killer cells, and 3.0L culture medium.

[0056] Using cytokines 200ng / ml interleukin-15, 100ng / ml interleukin-18 and 1000IU / ml interleukin-2 (both purchased from Miltenyi, Germany) were continuously cultured in serum-free medium AIM-V for 20 days to obtain 3.0×10 9 Natural killer...

Embodiment 2

[0058] Example 2 Preparation of cosmetic raw materials derived from human immune cells

[0059] Take the immune cells obtained in Example 1, add 10ml 1×PBS (pH=7.4) and mix well, then add 1ug protease inhibitor (Dalian Bao Biological Co., Ltd.), place it on ice and crush it in an ultrasonic breaker (Thermo Electric Corporation, USA) After 8 minutes, centrifuge at a high speed of 15,000 rpm for 20 minutes, and take the supernatant and add it to the culture medium in which the immune cells have been cultured.

[0060] The culture solution containing the immune cell lysate was added to a dialysis bag with a molecular weight of 500 (Spectrum, USA), and placed in a refrigerator for dialysis concentration, and the volume was concentrated by 5 times.

[0061] In addition, the culture solution containing the immune cell lysate can also be concentrated by ultrafiltration using a 500 molecular weight cellulose acetate membrane (Spectrum, Inc., USA), and concentrated 5 times in volume. ...

Embodiment 3

[0063] Example 3 Detection of cytokine expression and secretion levels in human immune cells

[0064]Take the immune cell culture fluid prepared in Example 1, the culture fluid containing the immune cell lysate and its concentrate, and use the ELISA kit (R&D Systems, Inc., USA) to detect the expression and secretion levels of the cells. A blank control was made with the AIM-V medium in which immune cells had not been cultured. Table 1 shows the expression and secretion levels of various cytokines in the culture solution of immune cells, the culture solution containing immune cell lysate and the concentrated solution, and the unit is ng / ml.

[0065] Table 1

[0066]

[0067] Table 1 shows that immune cell culture fluid, culture fluid containing lysate, and concentrated fluid secrete high levels of anti-inflammatory molecules and antioxidant molecules, and the cell lysate contains Nrf2 molecules with dual functions, indicating that these cytokines are involved in In terms o...

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Abstract

The invention provides a preparing method of immune-cell-derived biological-beautifying raw materials. The method includes the steps that a culturing solution for culturing immune cells and a protein suspension obtained by splitting immune cells are subjected to hyperconcentration to obtain immune-cell essence, wherein the essence is rich in anti-inflammatory and anti-oxidation ingredients and has the anti-inflammatory and anti-oxidation effects; the essence is further subjected to freezing drying to obtain freezing-dried powder which can be stored in a -80-DEG-C ultra-low-temperature refrigerator for a long time, and the anti-inflammatory and anti-oxidation skin biological-beautifying raw materials are obtained. When the biological-beautifying raw materials are added into face cream, skin cream and toner, anti-inflammatory and anti-oxidation face cream products can be prepared; when the biological-beautifying raw materials are added into masks, anti-inflammatory and anti-oxidation mask products can be prepared; the biological-beautifying raw materials can be directly prepared into anti-inflammatory and anti-oxidation essence products and can be used for treating skin acne, removing freckles, blocking the sun, resisting oxidation and the like.

Description

technical field [0001] The invention relates to a method for preparing immune cell-derived biocosmetic raw materials and the immune cell-derived biocosmetic product obtained by the method. In particular, the present invention relates to the use of protein suspension obtained from the culture medium of culturing immune cells and lysed immune cells, which is super-concentrated and prepared into immune cell essence, and then freeze-dried and prepared into lyophilized biological cosmetic raw materials, which are rich in Anti-inflammatory and anti-oxidant ingredients, with anti-inflammatory and anti-oxidative effects, can be prepared into products such as face cream, body lotion, toner, mask and essence, and can be used for skin acne, spot removal, sun protection and anti-oxidation and other beauty treatments. Background technique [0002] At present, all kinds of cosmetics are mainly based on chemical ingredients, and some beauty products prepared by adding some plant extracts o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/99A61Q19/00A61Q19/08A61Q19/02A61P17/02A61P17/10A61P17/18
CPCA61K8/99A61Q19/00A61Q19/02A61Q19/08
Inventor 陈建勋黄启明廖文敏
Owner ZICHENG RUISHENGHUI BEIJING BIOTECH DEV CO LTD
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