渗透压对伤口愈合相关细胞增殖的影响

发布时间：2018-06-10 20:10

本文选题：渗透压 + 细胞增殖　；参考：《兰州大学》2017年硕士论文

【摘要】：目的:探讨不同浓度的渗透压溶液对人胚皮肤成纤维细胞(Human embryonic skin fibroblast,HESF)、人永生化表皮细胞(Human immortalized epidermal cells,HIEC)、人脐静脉内皮细胞(Human umbilical vein endothelial cells,HUVEC)和表皮干细胞(Epidermal stem cells,ESC)增殖的影响。方法:利用中性蛋白酶-胰酶消化法及Ⅳ型胶原粘附法原代提取及培养ESC,免疫荧光进行鉴定。配制五种不同渗透压梯度DMEM培养基,冰点渗透压仪测其渗透压为:G1(低渗组:230mmol/L),G2(低渗组,262mmol/L),G3(对照组:293mmol/L),G4(高渗组:321mmol/L),G5(高渗组:354mmol/L)。HESF,HIEC,HUVEC分别在五组不同培养基中孵育1h、3h、6h、12h、24h、48h、及72h,倒置显微镜下观察其细胞形态,CCK-8试剂盒检测其不同时间点的细胞增殖率。结果:HESF在低渗和高渗培养条件下,细胞增殖率降低(P0.05)。HIEC在G1培养6、24、48h时,细胞增殖率降低(P0.05),12h时升高(P0.05);在G2中,1h、72h时细胞增殖率升高(P0.05),48h时降低(P0.05);在G4,G5培养条件下,细胞增殖率降低(P0.05)。HUVEC在低渗G1培养1、3、12和24h时,细胞增殖率降低(P0.05),在72h时细胞增殖率升高(P0.05);在G2、G4、G5条件下,细胞增殖率降低(P0.05)。ESC在G1培养6、24、48h时,细胞增殖率降低(P0.05);在G2中,48h、72h时,细胞增殖率降低(P0.05);在G4中,1h时细胞增殖率增高(P0.05),12h、24h、48h、72h时细胞增殖率降低(P0.05);在G5中,1h、3h、6h时细胞增殖率增高(P0.05),12h、48h、72h时细胞增殖率降低(P0.05)。结论:培养基渗透压梯度显著影响HESF、HIEC、HUVEC、ESC的增殖能力,且不同作用时间对HIEC,HUVEC的增殖能力的影响存在差异。通过本实验证实了渗透压对细胞增殖活力的影响,为临床调整创面渗透压促进伤口愈合以及组织工程技术利用渗透压培养表皮干细胞提供了理论依据。
[Abstract]:Objective: To investigate Human embryonic skin fibroblast (HESF), human immortalized epidermal cells (Human immortalized epidermal cells, HIEC), human umbilical vein endothelial cells (Human umbilical) and epidermal stem cells (epidermal stem cells) in different concentrations of osmotic pressure. Methods: using the neutral protease trypsin digestion method and the type IV collagen adhesion method to extract and culture ESC and identify the immunofluorescence. Five different osmotic pressure gradient DMEM medium were prepared. The osmotic pressure of ice point osmotic pressure instrument was used to measure the osmotic pressure of G1 (low permeability group: 230mmol/L), G2 (low permeability group, 262mmol/L), G3 (control group: 293mmol/L), G4 (32) 1mmol/L), G5 (hypertonic group: 354mmol/L).HESF, HIEC, HUVEC incubated 1H, 3h, 6h, 12h, 24h, 48h, and HUVEC respectively in the five different groups of cultures, and observed the cell morphology under the inverted microscope. In 6,24,48h, cell proliferation rate decreased (P0.05) and 12h increased (P0.05); in G2, 1H, 72h increased the cell proliferation rate (P0.05) and 48h decreased (P0.05). In 5 conditions, cell proliferation rate decreased (P0.05).ESC in G1 culture 6,24,48h, cell proliferation rate decreased (P0.05); in G2, 48h, 72h, cell proliferation rate decreased (P0.05); in G4, 1H cell proliferation rate increased (P0.05) The cell proliferation rate decreased (P0.05). Conclusion: the osmotic pressure gradient in culture medium significantly affects the proliferation of HESF, HIEC, HUVEC and ESC, and the effects of different time on the proliferation of HIEC and HUVEC are different. The effect of osmotic pressure on cell proliferation activity is confirmed by this experiment, and the wound healing and group of wound osmotic pressure are adjusted to promote wound healing and group. Textile engineering technology provides a theoretical basis for using osmotic pressure to culture epidermal stem cells.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R641

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