CN104997708A - Obtaining method of stem cell conditioned medium, and application of stem cell conditioned medium in skin ageing resistance - Google Patents

Obtaining method of stem cell conditioned medium, and application of stem cell conditioned medium in skin ageing resistance Download PDF

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Publication number
CN104997708A
CN104997708A CN201510397108.3A CN201510397108A CN104997708A CN 104997708 A CN104997708 A CN 104997708A CN 201510397108 A CN201510397108 A CN 201510397108A CN 104997708 A CN104997708 A CN 104997708A
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China
Prior art keywords
stem cell
conditioned medium
cell conditioned
medium
cells
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CN201510397108.3A
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Chinese (zh)
Inventor
郭子宽
赵靖
王娟
尹珊
李乔丽
芦俊萍
刘瑾
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Henan Zhong Ke Stem Cell Gene Engineering Co Ltd
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Henan Zhong Ke Stem Cell Gene Engineering Co Ltd
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Priority to CN201510397108.3A priority Critical patent/CN104997708A/en
Publication of CN104997708A publication Critical patent/CN104997708A/en
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Abstract

The invention belongs to the field of use of a stem cell conditioned medium in medicinal cosmetics, and especially relates to an application of the conditioned medium in skin ageing resistance. Researches find that a large amount of complex active substances secreted by stem cells are stored in a medium in the in vivo culture process of mesenchymal stem cells, and is called as the stem cell conditioned medium, and an obtaining method of the medium comprises the following steps: carrying out digestive passage on p2 generation mesenchymal stem cells by using 0.05% pancreatin, inoculating 2*10<6> into a 150mm culture dish, and culturing to obtain p3 generation cells; and sucking the obtained culture supernatant when 80% of the p3 generation cells fuse, washing by using PBS twice, adding a new serum-free medium, putting the obtained cells in a 37DEG C 5% CO2 incubator, culturing for 48h, and collecting the obtained culture supernatant to obtain the stem cell conditioned medium. The stem cell conditioned medium obtained in the invention can supplement skin cell metabolism promotion substances lacked in skins and also can induce new collagen synthesis in order to maintain elasticity of skins and reduce wrinkles.

Description

The acquisition of stem cell conditioned medium and the application in anti-skin aging
Technical field
The invention belongs to stem cell conditioned medium and apply to pharmaceutical cosmetic sector, the especially application of conditioned medium in anti-skin aging.
Background technology
Cosmeceutical is the complexing agent of cosmetics and medicinal physiologic therapy material.Along with population increases, the improvement of people living condition, with age, the collagen in application on human skin, elastin laminin, fibroblast and extracellular matrix protein thereof reduce gradually, cutis laxa, occur linear wrinkle and darker fold.Skin aging problem comprises elasticity loss, facial wrinkles, pigmentation, xerosis cutis and skin texture irregularity etc., and the topic of not still numerous ordinary people care, also becomes the content of scientists study.
How to use medicine to repair photo damaged skin, recover the content of skin elasticity, by the medical journal New England Journal of Medicine published in top level.
Plant stem cell extract is applied to clinical reduce wrinkle, obtain good effect, and it alleviates Pigmented function, points out the probability applied in beautifying face.
The Mechanism Study of human stem cell treatment wrinkle of skin, applies for science forefront result of study and has enriched hope in anti-skin aging.
Korea S carries out stem cell to improve looks one of maximum country in the world, use cell comprises endotheliocyte, mescenchymal stem cell and the adipose cell etc. that induced multi-potent stem cells (induced pluripotent stem cells, iPS cell) produces after differentiation-inducing.
IPS cell has been built, and uses iPS cell not relate to the medical ethics problems caused because drawing materials (bone marrow, umbilical cord and Placenta Hominis), but, due to iPS cell tumorigenesis risk, can only conditioned medium be used.
But mostly the effective active composition in stem cell conditioned medium is macromolecular substances, and keratodermatitis seriously hinders these material absorbings to enter skin histology.Therefore, how the material of stem cell secretion being imported skin, is the problem that stem cell conditioned medium reduce wrinkle need solve.
Summary of the invention
Intradermal injection mescenchymal stem cell, cell can be survived 1 year in topical dermal and hypodermic layer, and can not migrate to other Organ and tissue.Stem cell not only directly can fill wrinkle, by being divided into endotheliocyte like cell, promote revascularization, also by secretion activity material, the hyper-proliferative of check melanin cell, accelerate skin renewal speed, but find after deliberation to cultivate in vitro in mescenchymal stem cell process, these active substances storage secreted by cell is stayed in culture medium, be referred to as stem cell conditioned medium (stem cellconditioned medium), comprise the somatomedin secreted by cell and cytokine.
The acquisition of stem cell conditioned medium: p2 is gone down to posterity for mescenchymal stem cell 0.05% trypsinization, with 2 × 10 6p3 is for cell in the individual 150mm of being inoculated in culture dish; When p3 is for cell fusion 80%, suck culture supernatant, wash twice with PBS, add new serum-free medium, put into 37 DEG C, after cultivating 48h in the incubator of 5%CO2, collect culture supernatant and be stem cell conditioned medium.
The stem cell conditioned medium of gained in the present invention, both can supplement the material of the promotion skin cell metabolism lacked in skin, can induce newborn collage synthesis again, thus maintained skin elasticity, alleviated depth of folding.
Detailed description of the invention
Be described in detail below in conjunction with the content of embodiment to invention
Embodiment 1
Utilize micropin that stem cell conditioned medium is imported wrinkle position
Utilize micropin that stem cell conditioned medium is imported wrinkle position, once in a week, continuous 5 weeks.
Do dermal pathology analysis, after finding stem-cell therapy, skin collagen increases, and wrinkle shoals, and elasticity increases.
Micropin can obtain good effect in conjunction with stem cell conditioned medium.
Embodiment 2
Utilize segmented radio frequency in conjunction with micropin puncture technique, stem cell conditioned medium is imported gauffer position
Segmented radio frequency also obtain good therapeutic effect in conjunction with micropin puncture technique.
Do dermal pathology analysis, after finding stem-cell therapy, skin collagen increases, and wrinkle shoals, and elasticity increases.
Embodiment 3
Iontophoretic technology also can import skin histology by stem cell conditioned medium, and anti-wrinkle effect is good.
Do dermal pathology analysis, after finding stem-cell therapy, skin collagen increases, and wrinkle shoals, and elasticity increases.
Experiment proves the stem cell being derived from people's umbilical cord and Placenta Hominis, not only can produce the extracellular matrix such as collagen and fibronectin, also cytokine profiles and somatomedin can be secreted, stem cell conditioned medium contains somatomedin and cytokine isoreactivity material can activate skin flbroblast and horn cell, promote angiogenesis, induction collage synthesis, suppresses collagen degradation, thus repairs the skin of chronic injury.

Claims (1)

1. the application of stem cell conditioned medium in anti-skin aging.
The preparation method of the stem cell conditioned medium according to claim 1: p2 is gone down to posterity for mescenchymal stem cell 0.05% trypsinization, with 2 × 10 6p3 is for cell in the individual 150mm of being inoculated in culture dish; When p3 is for cell fusion 80%, suck culture supernatant, wash twice with PBS, add new serum-free medium, put into 37 DEG C, after cultivating 48h in the incubator of 5%CO2, collect culture supernatant and be stem cell conditioned medium.
CN201510397108.3A 2015-07-07 2015-07-07 Obtaining method of stem cell conditioned medium, and application of stem cell conditioned medium in skin ageing resistance Pending CN104997708A (en)

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CN201510397108.3A CN104997708A (en) 2015-07-07 2015-07-07 Obtaining method of stem cell conditioned medium, and application of stem cell conditioned medium in skin ageing resistance

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CN105238749A (en) * 2015-11-17 2016-01-13 广州赛莱拉干细胞科技股份有限公司 Method for resuscitating mesenchymal stem cells
CN105238750A (en) * 2015-11-17 2016-01-13 广州赛莱拉干细胞科技股份有限公司 Method for resuscitating umbilical cord mesenchymal stem cells
CN105687084A (en) * 2016-03-10 2016-06-22 广州赛莱拉干细胞科技股份有限公司 Skin-whitening and moisturizing toner and preparation method thereof
CN107661569A (en) * 2016-07-27 2018-02-06 上海坤爱生物科技股份有限公司 One kind is used for the oxidation resistant stem cell processed material micropin of skin
CN108373995A (en) * 2018-02-14 2018-08-07 浙江生创精准医疗科技有限公司 A kind of stem cell conditioned medium, preparation method and use
CN111454889A (en) * 2019-01-21 2020-07-28 芯芮生技开发股份有限公司 Application of stem cell anti-blue light damage efficacy composition
CN111454898A (en) * 2020-04-23 2020-07-28 中科晟和干细胞技术有限公司 Preparation method of mesenchymal stem cell culture medium for cosmetics
CN111511901A (en) * 2017-12-06 2020-08-07 李廷福 Culture medium composition for cell proliferation, skin regeneration and wrinkle improvement comprising blastodermal pluripotent stem cells isolated from avian eggs, neural stem cells, embryonic fibroblast conditioned medium as an effective ingredient
CN112220734A (en) * 2020-11-16 2021-01-15 郑州源创吉因实业有限公司 Skin-care spray containing stem cell active factors and preparation method thereof

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Publication number Priority date Publication date Assignee Title
CN105238749B (en) * 2015-11-17 2019-03-01 广州赛莱拉干细胞科技股份有限公司 A kind of method of recovery mesenchymal stem cell
CN105238750A (en) * 2015-11-17 2016-01-13 广州赛莱拉干细胞科技股份有限公司 Method for resuscitating umbilical cord mesenchymal stem cells
CN105238749A (en) * 2015-11-17 2016-01-13 广州赛莱拉干细胞科技股份有限公司 Method for resuscitating mesenchymal stem cells
CN105238750B (en) * 2015-11-17 2019-03-01 广州赛莱拉干细胞科技股份有限公司 A kind of method of recovery umbilical cord mesenchymal stem cells
CN105687084A (en) * 2016-03-10 2016-06-22 广州赛莱拉干细胞科技股份有限公司 Skin-whitening and moisturizing toner and preparation method thereof
CN105687084B (en) * 2016-03-10 2018-09-21 广州赛莱拉干细胞科技股份有限公司 A kind of skin whitening, moisturizing toner and preparation method thereof
CN107661569A (en) * 2016-07-27 2018-02-06 上海坤爱生物科技股份有限公司 One kind is used for the oxidation resistant stem cell processed material micropin of skin
CN111511901A (en) * 2017-12-06 2020-08-07 李廷福 Culture medium composition for cell proliferation, skin regeneration and wrinkle improvement comprising blastodermal pluripotent stem cells isolated from avian eggs, neural stem cells, embryonic fibroblast conditioned medium as an effective ingredient
CN108373995A (en) * 2018-02-14 2018-08-07 浙江生创精准医疗科技有限公司 A kind of stem cell conditioned medium, preparation method and use
CN108373995B (en) * 2018-02-14 2020-11-17 浙江生创精准医疗科技有限公司 Stem cell conditioned medium, preparation method and application thereof
CN111454889A (en) * 2019-01-21 2020-07-28 芯芮生技开发股份有限公司 Application of stem cell anti-blue light damage efficacy composition
CN111454898A (en) * 2020-04-23 2020-07-28 中科晟和干细胞技术有限公司 Preparation method of mesenchymal stem cell culture medium for cosmetics
CN112220734A (en) * 2020-11-16 2021-01-15 郑州源创吉因实业有限公司 Skin-care spray containing stem cell active factors and preparation method thereof

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Application publication date: 20151028