A mesenchymal stem cell epidermal differentiation inducer and its application method
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A differentiation inducer and stem cell technology, applied in the field of stem cell induction and differentiation, can solve problems such as low differentiation yield, and achieve the effects of strong cell activity, improved induction and differentiation efficiency, and high safety.
Active Publication Date: 2018-07-27
QINGDAO RESTORE BIOTECHNOLOGY CO LTD
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For example, some researchers used skin tissue homogenate as conditioned medium and added epidermal growth factor (EGF) to induce epidermal differentiation of stem cells, and proved that MSCs can also differentiate into epidermal cells in vitro by detecting the markers CK19 and CK14 of epidermal stem cells. However, the differentiation rate of this method is very low (the positive rate of CK19 is about 10%, and the positive rate of CK14 is about 12%)
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Embodiment 1
[0018] Embodiment 1 An agent for inducing epidermal differentiation of mesenchymal stem cells, characterized in that: the inducing agent is composed of KGF, BMP-4, PDGF-AB, VEGF, IL-2, and Forskolin.
Embodiment 2
[0019] Example 2 Mesenchymal stem cell epidermal differentiation inducer, the following raw materials were dissolved according to their respective characteristics, and human mesenchymal stem cell serum-free medium (LONZA, product number 00190632) was used as the substrate, and the content of each component was adjusted to the following volume: 1000ml: KGF (Sigma, K1751-10μg) 20μg, BMP-4 (Sigma, SRP3016-10μg) 5μg, PDGF-AB (PeproTech, 96-100-00AB-10μg) 5μg, VEGF (PeproTech, 96-100-20- 10 μg) 10 μg, IL-2 (PeproTech, 96-200-02-50 μg) 5 μg, Forskolin (Abcam, ab120058-100mg) 1 mg, mix well and filter to sterilize.
Embodiment 3
[0020] Example 3 Mesenchymal stem cell epidermal differentiation inducer, the following raw materials are dissolved according to their respective characteristics, and DMEM / F12+10%FBS is used as the matrix, and the content of each component is adjusted to 1000ml as follows: KGF (Sigma, K1751 -10 μg) 30 μg, BMP-4 (Sigma, SRP3016-10 μg) 10 μg, PDGF-AB (PeproTech, 96-100-00AB-10 μg) 10 μg, VEGF (PeproTech, 96-100-20-10 μg) 20 μg, IL-2 (PeproTech, 96-200-02-50μg) 10μg, Forskolin (Abcam, ab120058-100mg) 2mg, mix well and filter to sterilize.
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Abstract
The invention belongs to the technical field of induction and differentiation of stem cells, and in particular relates to an inducer of directed differentiation of mesenchymal stem cells. A mesenchymal stem cell epidermal differentiation inducer, which is composed of KGF, BMP‑4, PDGF‑AB, VEGF, IL‑2, and Forskolin. After adding complete medium, the concentration of each component is KGF 20~30μg / L , BMP-4 5-10 μg / L, PDGF-AB 5-10 μg / L, VEGF 10-20 μg / L, IL-2 5-10 μg / L, Forskolin 1-2 mg / L. The mesenchymal stem cell epidermal differentiation inducer of the present invention uses cytokines combined with small molecule compounds to efficiently induce mesenchymal stem cells to differentiate into epidermal cells, greatly improves the differentiation efficiency, and has strong cell activity after induction without cell transfection. Therefore, there is no genetic change and risk of cancer, no ethical issues, and high safety.
Description
technical field [0001] The invention belongs to the technical field of induction and differentiation of stem cells, and in particular relates to an inducer of directed differentiation of mesenchymal stem cells. Background technique [0002] The skin is the largest organ in the human body, accounting for about 16% of the body weight, and is mainly composed of epidermis, dermis and subcutaneous tissue. Clinically, due to skin damage caused by severe burns, chronic ulcers, trauma, etc., it is necessary to promote wound healing as soon as possible to minimize dysfunction caused by paralyzed scars. The development of tissue engineering technology and regenerative medicine has enabled the development of new skin regeneration products. The basic principle of tissue engineering is to culture and amplify seed cells in vitro, integrate them on a pre-designed biological scaffold to form a cell-scaffold complex, and then implant them into the corresponding lesion parts of the body to r...
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