Medicine for accelerating skin repair and regeneration, preparation method thereof and application thereof
A drug and skin technology, applied in the field of drugs for accelerating skin layer repair and skin accessory organ regeneration
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Embodiment 1
[0042] Example 1: Preparation method, optimization of preparation conditions and identification of epidermal stem cells
[0043] 1. Preparation method of epidermal stem cells
[0044] At present, the preparation method of epidermal stem cells mainly includes two steps: 1) Firstly, a single skin epidermis is obtained through the functional separation of the junction between the epidermis and the dermis by neutral protease, which avoids the pollution of dermal cells; and, because trypsin damages the cells Therefore, the neutral protease digestion method is firstly used to separate the epidermal layer of the skin by using its function of hydrolyzing the junction between the epidermis and the dermis, and then the epidermal layer of the skin is digested with trypsin to obtain a single cell. It is beneficial to the protection of the activity of the digested cells and ensures that a sufficient number of active target cells are obtained; 2) the epidermal stem cells are then separated ...
Embodiment 2
[0099] Example 2: Preparation of bone marrow mesenchymal stem cells
[0100] The preparation of bone marrow mesenchymal stem cells can refer to the literature (Cai-Xia He, Ni Li, Yu-Lan Hu, Xiu-Mei Zhu, Hai-Jie Li, Min Han, Pei-Hong Miao, Zhong-Jie Hu, Gang Wang, Wen-Quan Liang, Yasuhiko Tabata, Jian-Qing Gao*, Effective gene delivery to mesenchymal stem cells based on the novel reverse transfection and three-dimensional cell culture system, Pharm Res, 2011, 28(7): 1577-1590)
[0101] 1) Preparation and in vitro culture of primary rat bone marrow mesenchymal stem cells (BMSCs): 3-week-old SD rats were sacrificed by neck pulling, the tibia and femur were separated, and the containing The tibia and femur were repeatedly washed with serum culture solution, and the obtained bone marrow was passed through a 200-mesh cell sieve (Shanghai Ruigu Biotechnology Co., Ltd.), centrifuged at 1000 rpm for 5 minutes, and the supernatant was discarded. After centrifugation, add the DMEM low-s...
Embodiment 3
[0104] Embodiment 3: the preparation of the medicine that accelerates skin layer repair and skin accessory organ regeneration
[0105] 1) The epidermal stem cells (ESCs) prepared by experimental group 7# in Example 1 were first washed twice with PBS, and then washed with trypsin-ethylenediaminetetraacetic acid (Trypsin-EDTA) containing 0.05% by weight of pancreatic Enzyme cell digestion solution (trypsin cell digestion solution obtained by dissolving trypsin-ethylenediaminetetraacetic acid in phosphate buffer at pH = 7.4) was digested for 7 minutes, and then added to the epidermal stem cells digested by trypsin cell digestion solution containing The DMEM low-sugar / F12 culture fluid of fetal bovine serum and penicillin streptomycin (the DMEM low-sugar / F12 culture fluid that contains fetal bovine serum and penicillin streptomycin is made of 10% fetal bovine serum by weight percentage, and the weight percentage is 1% penicillin, streptomycin, DMEM low-sugar culture medium of 59.3...
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