本文選題：滲透壓 + 細胞增殖��； 參考：《蘭州大學》2017年碩士論文

【摘要】：目的:探討不同濃度的滲透壓溶液對人胚皮膚成纖維細胞(Human embryonic skin fibroblast,HESF)、人永生化表皮細胞(Human immortalized epidermal cells,HIEC)、人臍靜脈內皮細胞(Human umbilical vein endothelial cells,HUVEC)和表皮干細胞(Epidermal stem cells,ESC)增殖的影響。方法:利用中性蛋白酶-胰酶消化法及Ⅳ型膠原粘附法原代提取及培養(yǎng)ESC,免疫熒光進行鑒定。配制五種不同滲透壓梯度DMEM培養(yǎng)基,冰點滲透壓儀測其滲透壓為:G1(低滲組:230mmol/L),G2(低滲組,262mmol/L),G3(對照組:293mmol/L),G4(高滲組:321mmol/L),G5(高滲組:354mmol/L)。HESF,HIEC,HUVEC分別在五組不同培養(yǎng)基中孵育1h、3h、6h、12h、24h、48h、及72h,倒置顯微鏡下觀察其細胞形態(tài),CCK-8試劑盒檢測其不同時間點的細胞增殖率。結果:HESF在低滲和高滲培養(yǎng)條件下,細胞增殖率降低(P0.05)。HIEC在G1培養(yǎng)6、24、48h時,細胞增殖率降低(P0.05),12h時升高(P0.05);在G2中,1h、72h時細胞增殖率升高(P0.05),48h時降低(P0.05);在G4,G5培養(yǎng)條件下,細胞增殖率降低(P0.05)。HUVEC在低滲G1培養(yǎng)1、3、12和24h時,細胞增殖率降低(P0.05),在72h時細胞增殖率升高(P0.05);在G2、G4、G5條件下,細胞增殖率降低(P0.05)。ESC在G1培養(yǎng)6、24、48h時,細胞增殖率降低(P0.05);在G2中,48h、72h時,細胞增殖率降低(P0.05);在G4中,1h時細胞增殖率增高(P0.05),12h、24h、48h、72h時細胞增殖率降低(P0.05);在G5中,1h、3h、6h時細胞增殖率增高(P0.05),12h、48h、72h時細胞增殖率降低(P0.05)。結論:培養(yǎng)基滲透壓梯度顯著影響HESF、HIEC、HUVEC、ESC的增殖能力,且不同作用時間對HIEC,HUVEC的增殖能力的影響存在差異。通過本實驗證實了滲透壓對細胞增殖活力的影響,為臨床調整創(chuàng)面滲透壓促進傷口愈合以及組織工程技術利用滲透壓培養(yǎng)表皮干細胞提供了理論依據。
[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.
【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R641

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