肌腱细胞与聚二甲基硅氧烷多孔材料复合培养后的玻璃化低温保存
发布时间：2018-07-29 14:13
【摘要】：背景:大量实验已证明,玻璃化低温保存能获得更高的细胞存活率。目的:观察玻璃化低温保存对肌腱细胞与聚二甲基硅氧烷材料复合物的影响。方法:取共培养9-14 d的肌腱细胞-聚二甲基硅氧烷复合物,分别以10%二甲亚砜、玻璃化低温保存液VS55、21%二甲亚砜进行低温保存和复苏。复苏培养1 h后,进行死/活双色荧光染色和流式细胞技术分析,观察肌腱细胞存活率,以新鲜培养的肌腱细胞-聚二甲基硅氧烷复合物为对照。结果与结论:1死/活双色荧光染色:10%二甲亚砜组肌腱细胞从聚二甲基硅氧烷支架材料孔隙表面上剥脱,呈双染的不规则细胞形态;VS55组和21%二甲亚砜组存在单染绿色荧光的梭形和球形细胞,也存在红绿双染的不规则形态细胞,两组细胞观测密度较对照组明显下降;2流式细胞技术分析:对照组细胞大小均匀;10%二甲亚砜组由于细胞量少而不足以进行流式细胞检测;VS55组以较小体积的细胞颗粒为主;21%二甲亚砜组细胞体积大小与新鲜对照组类似,同时存在较小颗粒;3细胞回收率与存活率:VS55组细胞相对存活率低于21%二甲亚砜组(64.9%,76.2%,P0.05);10%二甲亚砜不能获取足够细胞,未能行细胞计数;4结果表明:采用21%二甲亚砜玻璃化低温保存有利于提高细胞的存活率,保持肌腱细胞-支架结合完整。
[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.
【作者单位】： 四川大学华西医院生物治疗国家重点实验室;西南医科大学附属医院骨与关节外科;德阳市人民医院;成都市第一人民医院;
【分类号】：R318
本文编号：2152937
[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.
【作者单位】： 四川大学华西医院生物治疗国家重点实验室;西南医科大学附属医院骨与关节外科;德阳市人民医院;成都市第一人民医院;
【分类号】：R318
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1 王治;玻璃化低温保存肌腱细胞与PDMS材料复合物的实验研究[D];四川大学;2007年
，本文编号：2152937
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