【摘要】：冷凍干燥是長(zhǎng)期有效保存細(xì)胞的手段之一。本文以肝癌細(xì)胞Hep-G2為研究對(duì)象,選取了不同體積分?jǐn)?shù)的Me2SO、丙三醇及不同質(zhì)量濃度的PVP、復(fù)方保護(hù)劑進(jìn)行冷凍干燥保存,篩選出最佳保護(hù)劑配方。此外,將細(xì)胞在不同摩爾濃度的海藻糖溶液中孵育,通過(guò)檢測(cè)細(xì)胞回收率、存活率和24 h貼壁率,探究胞內(nèi)海藻糖對(duì)細(xì)胞冷凍干燥的影響。結(jié)果表明:添加40%PVP(w/v)+10%甘油(v/v)+15%FBS(v/v)+20%海藻糖(w/v)保護(hù)劑的細(xì)胞,在復(fù)水后回收率、存活率和24 h貼壁率分別為29.58%、42.18%和18.71%,與對(duì)照組相比三項(xiàng)指標(biāo)均得到有效提高,該種保護(hù)劑的效果最佳;當(dāng)胞外海藻糖摩爾濃度為800 mmol/L時(shí),凍干效果最好,肝癌細(xì)胞Hep-G2在復(fù)水后回收率、存活率和24 h貼壁率分別為27.81%,66.65%和33.68%,存活率和貼壁率顯著高于其他組。
[Abstract]:Freeze-drying is one of the means of long-term preservation of cells. In this paper, we select different volume fractions of Me2SO, glycerol and different mass concentrations of PVP and compound protectants for freeze-drying and storage, and screen out the optimal formulation of protective agent. In addition, the cells were incubated in a different molar concentration of trehalose solution, and the effect of intracellular trehalose on the freeze-drying of the cells was explored by the detection of cell recovery, survival and 24 h adherence. The results showed that 40% PVP (w/ v) + 10% glycerol (v/ v) + 15% FBS (v/ v) + 20% trehalose (w/ v) protectants were added. The recovery rate, survival rate and the adherence rate of 24 h were 29. 58%, 42. 18% and 18. 71%, respectively. When the molar concentration of the extracellular trehalose is 800 mmol/ L, the freeze-drying effect is the best, and the recovery rate, the survival rate and the 24-hour adherence rate of the Hep-G2 in the liver cancer cell are 27. 81%, 66. 65% and 33. 68%, respectively, and the survival rate and the adherence rate are significantly higher than the other groups.
【作者單位】： 上海理工大學(xué)醫(yī)療器械與食品學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(51376132)資助項(xiàng)目~~
【分類(lèi)號(hào)】：R73-3
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本文編號(hào)：2361106