Tissue engineered skin with basilar membrane and construction method thereof
A tissue-engineered skin and construction method technology, applied in artificial cell constructs, medical science, prostheses, etc., to achieve the effect of rapid epidermal growth, wide source of materials, and low cost
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Embodiment 1
[0024] Example 1. Preparation of Active Skin Substitutes Containing Basement Membrane
[0025] 1. Isolation and culture of human keratinocytes and fibroblasts
[0026] The discarded foreskin tissue after circumcision was used to separate and culture keratinocytes and fibroblasts by enzymatic digestion. Separately prepared into a single cell suspension, according to 2-3 × 10 5 Cells per ml were seeded in culture flasks and placed in a 37°C incubator. Keratinocytes were cultured in serum-free medium (DK-SFM, Gibco, USA), and fibroblasts were cultured in complete DMEM medium (Gibco, USA). USA) for culture, and when the cells reach 70%-80% confluence, passage and expansion are carried out.
[0027] 2. Surface modified modified amniotic membrane
[0028] Strip off the fresh amniotic membrane under aseptic conditions (taken from the placental tissue of caesarean section women who are negative for hepatitis virus antibody, syphilis antibody and HIV), bluntly separate the chorion, ...
Embodiment 2
[0031] Embodiment 2. Animal transplantation test of tissue engineered skin of the present invention
[0032] Male Balb / c-nu mice (nude mice, provided by Shanghai Xipuer-Bikay Experimental Animal Co., Ltd.), body weight 18 ± 2 grams, 16, were divided into two groups, respectively, the tissue engineered skin containing amniotic membrane of the present invention Group A and the traditional active skin substitute without amniotic membrane Group B: purely constructed with fibrin as a matrix (see A.L.Mazlyzama, B.S.Aminuddinb, et al.Reconstruction of living bilayerhuman skin equivalent utilizing human fibrin as a scaffold.Burns, 2007, 33(3): 355-363).
[0033]After the nude mice were anesthetized with ketamine intraperitoneally, the skin and hair on the back were shaved, and the full-thickness skin on the ventral side of the spine and the deep fascia up to the muscle layer were excised. Group A: The tissue-engineered skin cultured in vitro for 2-3 weeks described in Example 1 was t...
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