CN113197920A - Preparation method of embryonic stem cell culture extract for skin aging resistance - Google Patents
Preparation method of embryonic stem cell culture extract for skin aging resistance Download PDFInfo
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- CN113197920A CN113197920A CN202110546508.1A CN202110546508A CN113197920A CN 113197920 A CN113197920 A CN 113197920A CN 202110546508 A CN202110546508 A CN 202110546508A CN 113197920 A CN113197920 A CN 113197920A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/54—Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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Abstract
The invention discloses a preparation method of an embryonic stem cell culture extract for skin aging resistance, which relates to the technical field of cosmetology and comprises the following steps: s1: inoculating the embryonic stem cells to an amplification culture medium for culturing, collecting culture supernatant of the embryonic stem cells, and extracting cell secretions in the culture supernatant; s2: concentrating and filtering the extracted culture secretion to obtain the embryonic stem cell culture extract. According to the preparation method of the embryonic stem cell culture extract for skin aging resistance, human embryonic stem cells are preferably used as a source of the culture extract. The culture supernatant of the embryonic stem cells is rich in active factor components such as epidermal growth factors, vascular endothelial growth factors, stem cell factors and the like, the factors can promote the growth and proliferation of stem cells in vivo and promote fibroblasts and the like in the skin to secrete more collagen, and the biological effects have the advantages which are incomparable with the traditional cosmetics and can obviously relieve the aging of the skin.
Description
Technical Field
The invention relates to the technical field of beauty treatment, in particular to a preparation method of an embryonic stem cell culture extract for skin aging resistance.
Background
Embryonic Stem Cells (ESCs) are a class of cells isolated from early embryos, have the characteristics of in vitro culture of unlimited proliferation, self-renewal and multidirectional differentiation, can be differentiated into any cell of three germ layers, have the capacity of developing into various tissues and participate in the formation of partial tissues.
The embryonic stem cells can secrete a plurality of factors in the culture process, mainly comprise factors related to pluripotency, transcription factors, mRNA, microRNA and the like, the factors can inhibit the cell aging process, change the epigenetic characteristics of target cells, reverse effector cells to a low differentiation state, play a role similar to tissue stem cells, such as the functions of repairing blood vessels, inhibiting inflammation, promoting the proliferation and differentiation of the tissue stem cells and the like, and play an important role in injury repair and tissue regeneration.
Most of cosmetic preparations on the market have the problems of single active ingredient, low individual pertinence, poor product effect, serious product homogenization and the like. The factors secreted by the embryonic stem cells can provide various nutrient and substance information required by cell growth and differentiation, can quickly activate a human body repair system, and can carry out intelligent repair according to different skin problems. Therefore, the development of a cosmetic formulation capable of surpassing the optimal repair efficacy of a single cytokine is a problem to be solved.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a preparation method of a culture extract of embryonic stem cells for skin aging resistance, which solves the problems in the background art.
In order to achieve the purpose, the invention is realized by the following technical scheme: a method for preparing an extract of embryonic stem cells for skin aging resistance comprises the following steps:
s1: inoculating the embryonic stem cells to an amplification culture medium for culturing, collecting culture supernatant of the embryonic stem cells, and extracting cell secretions in the culture supernatant;
s2: concentrating and filtering the extracted culture secretion to obtain the embryonic stem cell culture extract.
Preferably, when the embryonic stem cells are cultured in S1, the cell culture plate is coated with matrigel.
Preferably, the stem cell culture supernatant stably cultured for 2 days is collected and concentrated in the step S2.
Preferably, in the step S2, the cell culture supernatant is concentrated by filtration through 0.45 μm and 0.22 μm filters.
An application method of an embryonic stem cell culture extract for skin aging resistance comprises the following steps:
s01: performing skin detection on the skin of a subject to obtain a skin data report of the subject;
s02: the subject uses the embryonic stem cell culture extract to carry out skin care, and 1mL of the extract is taken in the morning and at night every day and is externally applied on the surface layer of facial skin;
s03: after the skin test is carried out for 7 days and 30 days respectively, skin data reports of the testee are obtained, and the skin states before and after the use are compared.
The invention provides a preparation method of an embryonic stem cell culture extract for skin aging resistance, which has the following beneficial effects:
1. according to the preparation method of the embryonic stem cell culture extract for skin aging resistance, human embryonic stem cells are preferably used as a source of the culture extract. The culture supernatant of the embryonic stem cells is rich in active factor components such as epidermal growth factors, vascular endothelial growth factors, stem cell factors and the like, the factors can promote the growth and proliferation of stem cells in vivo and promote fibroblasts and the like in the skin to secrete more collagen, and the biological effects have the advantages which are incomparable with the traditional cosmetics and can obviously relieve the aging of the skin.
2. The preparation method of the anti-aging embryonic stem cell culture extract for skin adopts the serum-free culture medium to culture the embryonic stem cells, is safe and reliable, and can not cause immunological rejection reaction like the traditional serum culture method.
3. According to the preparation method of the embryonic stem cell culture extract for skin aging resistance, the embryonic stem cell culture solution is concentrated and purified by adopting tangential flow ultrafiltration, the purification efficiency is high, and the preparation method is suitable for large-scale production of the embryonic stem cell culture extract.
4. According to the preparation method of the embryonic stem cell culture extract for skin aging resistance, the prepared embryonic stem cell culture extract has remarkable beautifying and aging resistance effects, and a subject using the embryonic stem cell culture extract has the advantages of wrinkle disappearance, elasticity improvement, skin color whitening, pore reduction, smooth and tender skin and good lasting effect.
Drawings
FIG. 1 is a diagram illustrating the steps of preparing an extract of embryonic stem cell for skin aging prevention according to the present invention;
FIG. 2 is a diagram illustrating the steps of applying the cultured extract of embryonic stem cells for skin aging prevention according to the present invention;
FIG. 3 is a graph showing the improvement of facial skin before and after 30 persons of the present invention used embryonic stem cell culture extracts several times.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
Example one
Referring to fig. 1, the present invention provides a technical solution: a method for preparing an extract of embryonic stem cells for skin aging resistance comprises the following steps:
s1: inoculating the embryonic stem cells to an amplification culture medium for culturing, collecting culture supernatant of the embryonic stem cells, and extracting cell secretions in the culture supernatant;
s2: concentrating and filtering the extracted culture secretion to obtain the embryonic stem cell culture extract.
As will be understood by those skilled in the art:
1. cell culture supernatant pretreatment: embryonic stem cell culture supernatant (fresh or re-warmed at-20 ℃) was taken, centrifuged at 3000g for 15min to remove dead cells and debris, and the supernatant was transferred to a sterilized 1L blue-capped vial (not to be poured too clean).
2. Filtering to remove large particles: filtering the supernatant of step 1 with 0.45 μm and 0.22 μm water-based membranes, and soaking the filter membrane in 70% ethanol for sterilization before use. The 0.22 μm filtered supernatant was transferred to a blue-capped vial.
3. Tangential flow ultrafiltration concentration of cell supernatant:
1) putting a liquid inlet/return pipeline into the cell supernatant prepared in the step 2, putting a filtering pipeline into a 1L waste liquid bottle, starting a peristaltic pump and regulating the speed until the pressure indicates 2.5bar, and continuously circulating and concentrating until about 10-15 ml of concentrated solution is left;
2) the circulation speed is reduced to 20-40ml/min and the circulation system is continued for 1-2min to increase the sample recovery rate.
3) And taking out the liquid inlet pipe, pumping the residual samples in the system to the sample bottle, adding about 10ml of DPBS into the liquid inlet pipe, cleaning and pumping into the sample bottle.
4.0.22 μm membrane filtration: the concentrated solution was collected by filtration using millipore 0.22 μm filter to obtain embryonic stem cell culture extract, which was refrigerated at-80 ℃ for future use.
Further, when culturing the embryonic stem cells in S1, the cell culture plate is coated with matrigel.
As will be understood by those skilled in the art, the culture effect is made better.
Further, the culture supernatant of the stem cells stably cultured for 2 days was collected and concentrated in step S2.
As will be appreciated by those skilled in the art, the concentration of the stem cell culture supernatant is made to be effective.
Further, in the step S2, in the concentration of the cell culture supernatant, filtration was performed using 0.45 μm and 0.22 μm filters in this order to obtain a concentrated solution.
As will be appreciated by those skilled in the art, the resulting concentrate will contain fewer impurities.
Example two
Referring to fig. 2, an application method of the embryonic stem cell culture extract for skin aging resistance includes the following steps:
s01: performing skin detection on the skin of a subject to obtain a skin data report of the subject;
s02: the subject uses the embryonic stem cell culture extract to carry out skin care, and 1mL of the extract is taken in the morning and at night every day and is externally applied on the surface layer of facial skin;
s03: after the skin test is carried out for 7 days and 30 days respectively, skin data reports of the testee are obtained, and the skin states before and after the use are compared.
As will be understood by those skilled in the art:
1. stability test
The obtained embryonic stem cell culture extract is stored at-20 deg.C. After six months, no significant change was observed.
2. Validity test
Selecting 30 subjects, detecting the skin of the subjects (CBS-807 skin analysis system) to detect the elasticity, pore size, moisture, wrinkles and skin color of the skin to obtain skin reports of the subjects, trying the obtained embryonic stem cell culture extract by the subjects, applying the embryonic stem cell culture extract on the surface layer of the skin in an external application mode, cleaning the face in the morning and at night every day, then applying corresponding samples for 2 times, respectively applying 1mL each time, and respectively carrying out skin detection again after continuously using for 7 and 30 days. The results are shown in FIG. three.
As can be seen from fig. 3, the embryonic stem cell culture extract provided by the present invention has excellent anti-aging and cosmetic effects, can effectively improve the problems of facial skin elasticity, moisture, wrinkles, spots, large pores, etc., and has a repairing effect on facial skin. In addition, 5 subjects with wrinkles, spots, and rough skin were selected, and the embryonic stem cell extracts were applied to the face for 7 days after cleansing the face every morning and evening. In total, after 5 subjects used for 7 days continuously, the wrinkles on the face became light and the skin became white and more delicate and smooth. When the embryonic stem cell extract is used by male subjects, wrinkles around eyes disappear, skin color becomes white, and the skin is more delicate.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (5)
1. A preparation method of an embryonic stem cell culture extract for skin aging resistance is characterized by comprising the following steps:
s1: inoculating the embryonic stem cells to an amplification culture medium for culturing, collecting culture supernatant of the embryonic stem cells, and extracting cell secretions in the culture supernatant;
s2: concentrating and filtering the extracted culture secretion to obtain the embryonic stem cell culture extract.
2. The method for preparing the culture extract of embryonic stem cells for skin aging prevention according to claim 1, wherein the culture extract of embryonic stem cells for skin aging prevention comprises: when the S1 is used for culturing the embryonic stem cells, the cell culture plate is coated by matrigel.
3. The method for preparing the culture extract of embryonic stem cells for skin aging prevention according to claim 1, wherein the culture extract of embryonic stem cells for skin aging prevention comprises: in the step S2, a culture supernatant of the stem cells stably cultured for 2 days was collected and concentrated.
4. The method for preparing the culture extract of embryonic stem cells for skin aging prevention according to claim 1, wherein the culture extract of embryonic stem cells for skin aging prevention comprises: and in the step S2, filtering by using 0.45 mu m and 0.22 mu m filter membranes in sequence in the process of concentrating the cell culture supernatant to obtain a concentrated solution.
5. An application method of an embryonic stem cell culture extract for skin aging resistance is characterized by comprising the following steps:
s01: performing skin detection on the skin of a subject to obtain a skin data report of the subject;
s02: the subject uses the embryonic stem cell culture extract to carry out skin care, and 1mL of the extract is taken in the morning and at night every day and is externally applied on the surface layer of facial skin;
s03: after the skin test is carried out for 7 days and 30 days respectively, skin data reports of the testee are obtained, and the skin states before and after the use are compared.
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| CN202110546508.1A CN113197920A (en) | 2021-05-19 | 2021-05-19 | Preparation method of embryonic stem cell culture extract for skin aging resistance |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113952281A (en) * | 2021-11-08 | 2022-01-21 | 云南贺尔思细胞生物技术有限公司 | Antiallergic composition containing stem cell extract and preparation method thereof |
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| CN101497874A (en) * | 2009-03-04 | 2009-08-05 | 中山大学中山眼科中心 | Method for promoting somatic cell proliferation |
| CN102125514A (en) * | 2010-01-18 | 2011-07-20 | 上海众申生物科技有限公司 | Synergistic anti-ageing function of stem cells, non-depolarizing muscle relaxants and hyaluronic acid |
| CN105055265A (en) * | 2015-07-21 | 2015-11-18 | 北京博士园科技发展有限公司 | Product capable of maintaining beauty and keeping young and preparation method of product |
| CN109602766A (en) * | 2019-01-25 | 2019-04-12 | 汪泱 | A kind of application of the excretion body in human pluripotent stem cells source |
| CN110974770A (en) * | 2019-12-26 | 2020-04-10 | 苏州克劳丽化妆品有限公司 | Anti-aging composition, preparation and preparation method |
-
2021
- 2021-05-19 CN CN202110546508.1A patent/CN113197920A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101497874A (en) * | 2009-03-04 | 2009-08-05 | 中山大学中山眼科中心 | Method for promoting somatic cell proliferation |
| CN102125514A (en) * | 2010-01-18 | 2011-07-20 | 上海众申生物科技有限公司 | Synergistic anti-ageing function of stem cells, non-depolarizing muscle relaxants and hyaluronic acid |
| CN105055265A (en) * | 2015-07-21 | 2015-11-18 | 北京博士园科技发展有限公司 | Product capable of maintaining beauty and keeping young and preparation method of product |
| CN109602766A (en) * | 2019-01-25 | 2019-04-12 | 汪泱 | A kind of application of the excretion body in human pluripotent stem cells source |
| CN110974770A (en) * | 2019-12-26 | 2020-04-10 | 苏州克劳丽化妆品有限公司 | Anti-aging composition, preparation and preparation method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113952281A (en) * | 2021-11-08 | 2022-01-21 | 云南贺尔思细胞生物技术有限公司 | Antiallergic composition containing stem cell extract and preparation method thereof |
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Application publication date: 20210803 |