【摘要】：背景:大量實驗已證明,玻璃化低溫保存能獲得更高的細(xì)胞存活率。目的:觀察玻璃化低溫保存對肌腱細(xì)胞與聚二甲基硅氧烷材料復(fù)合物的影響。方法:取共培養(yǎng)9-14 d的肌腱細(xì)胞-聚二甲基硅氧烷復(fù)合物,分別以10%二甲亞砜、玻璃化低溫保存液VS55、21%二甲亞砜進(jìn)行低溫保存和復(fù)蘇。復(fù)蘇培養(yǎng)1 h后,進(jìn)行死/活雙色熒光染色和流式細(xì)胞技術(shù)分析,觀察肌腱細(xì)胞存活率,以新鮮培養(yǎng)的肌腱細(xì)胞-聚二甲基硅氧烷復(fù)合物為對照。結(jié)果與結(jié)論:1死/活雙色熒光染色:10%二甲亞砜組肌腱細(xì)胞從聚二甲基硅氧烷支架材料孔隙表面上剝脫,呈雙染的不規(guī)則細(xì)胞形態(tài);VS55組和21%二甲亞砜組存在單染綠色熒光的梭形和球形細(xì)胞,也存在紅綠雙染的不規(guī)則形態(tài)細(xì)胞,兩組細(xì)胞觀測密度較對照組明顯下降;2流式細(xì)胞技術(shù)分析:對照組細(xì)胞大小均勻;10%二甲亞砜組由于細(xì)胞量少而不足以進(jìn)行流式細(xì)胞檢測;VS55組以較小體積的細(xì)胞顆粒為主;21%二甲亞砜組細(xì)胞體積大小與新鮮對照組類似,同時存在較小顆粒;3細(xì)胞回收率與存活率:VS55組細(xì)胞相對存活率低于21%二甲亞砜組(64.9%,76.2%,P0.05);10%二甲亞砜不能獲取足夠細(xì)胞,未能行細(xì)胞計數(shù);4結(jié)果表明:采用21%二甲亞砜玻璃化低溫保存有利于提高細(xì)胞的存活率,保持肌腱細(xì)胞-支架結(jié)合完整。
[Abstract]:Background: a large number of experiments have shown that cryopreservation can achieve higher cell viability. Aim: to observe the effect of cryopreservation on the composite of tendon cells and polydimethylsiloxane. Methods: 10% dimethyl sulfoxide (DMSO) and VS55C21% dimethyl sulfoxide (DMSO) were used for cryopreservation and resuscitation of tendon cells and polydimethylsiloxane complex co-cultured for 9-14 days. After 1 hour of resuscitation culture, the survival rate of tendon cells was observed by fluorescence staining and flow cytometry. The fresh cultured tendon cells and polydimethylsiloxane complex were used as control. Results and conclusion the dead / live bichromatic fluorescence staining of 10% dimethyl sulfoxide group with 10% dimethyl sulfoxide removed the tendon cells from the pore surface of polydimethylsiloxane scaffolds. In group VS55 and group 21% dimethyl sulfoxide, there were fusiform and globular cells with green fluorescence and irregular cells with double staining of red and green. The cell density in both groups was significantly lower than that in the control group. Analysis of flow cytometry showed that the cell size of the control group was uniform and 10% dimethyl sulfoxide group was not enough to carry out flow cytometry to detect the cell size of VS55 group because of the small number of cells. The cell size of 21% dimethyl sulfoxide group was similar to that of fresh control group. At the same time, there was a smaller cell recovery and survival rate. The relative survival rate of the cells in the 10% dimethyl sulfoxide group was lower than that in the 21% dimethyl sulfoxide group (64.9% vs 76.22%), and 10% dimethyl sulfoxide could not obtain enough cells. The results showed that cryopreservation with 21% dimethyl sulfoxide vitrification could improve the cell survival rate and keep the tendon cell-scaffold intact.
【作者單位】： 四川大學(xué)華西醫(yī)院生物治療國家重點(diǎn)實驗室;西南醫(yī)科大學(xué)附屬醫(yī)院骨與關(guān)節(jié)外科;德陽市人民醫(yī)院;成都市第一人民醫(yī)院;
【分類號】：R318

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1 王治;玻璃化低溫保存肌腱細(xì)胞與PDMS材料復(fù)合物的實驗研究[D];四川大學(xué);2007年

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