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支架内连续介质流动结合软骨细胞接种和培养系统用于组织工程气管构建。

Intra-scaffold continuous medium flow combines chondrocyte seeding and culture systems for tissue engineered trachea construction.

作者信息

Tan Qiang, Hillinger Sven, van Blitterswijk Clemens A, Weder Walter

机构信息

Clinic of Thoracic Surgery, University Hospital Zurich, Zurich, Switzerland.

出版信息

Interact Cardiovasc Thorac Surg. 2009 Jan;8(1):27-30. doi: 10.1510/icvts.2008.179804. Epub 2008 Jun 12.

DOI:10.1510/icvts.2008.179804
PMID:18550604
Abstract

In this study we tested the possibility of seeding chondrocytes into poly (ethylene glycol)-terephthalate-poly (butylene terephthalate) PEOT/PBT scaffold through an intra-scaffold medium flow and the impact of this continuous medium flow on subsequent chondrocyte-scaffold culture. Eight cubic PEOT/PBT co-polymers (1 cm(3)) were assigned into two groups. In the semi-dynamic seeding group a continuous medium flow was created inside the scaffolds by a pump system. Around six million chondrocytes were harvested each day, suspended in 1 ml medium and delivered onto the scaffold through the perfusion for a sequential five days. Traditional chondrocytes directly seeding and static culture method was performed as control. Scanning electron microscopy (SEM) and histology assessments were performed to evaluate the distribution of chondrocytes inside the scaffolds and MTT test was chosen to check cell vitality. SEM pictures and histology slices from the perfusion group showed a better three-dimensional cell growth and extensive cell distribution inside the scaffolds; while in the control group chondrocytes only dispersedly formed a monolayer on the surface of scaffolds. Accordingly, MTT results from the perfusion group were much higher than those from control group (0.123 vs. 0.067, P<0.01). Continuous medium perfusion inside PEOT/PBT scaffold effectively combines chondrocyte seeding and culture systems for the reconstruction of tissue engineered trachea.

摘要

在本研究中,我们测试了通过支架内的介质流动将软骨细胞接种到聚(乙二醇)-对苯二甲酸酯-聚(丁二醇对苯二甲酸酯)(PEOT/PBT)支架中的可能性,以及这种连续介质流动对后续软骨细胞-支架培养的影响。将八个立方的PEOT/PBT共聚物(1立方厘米)分为两组。在半动态接种组中,通过泵系统在支架内部产生连续的介质流动。每天收获约六百万个软骨细胞,悬浮于1毫升培养基中,并通过灌注在连续五天的时间里输送到支架上。采用传统的软骨细胞直接接种和静态培养方法作为对照。进行扫描电子显微镜(SEM)和组织学评估以评价软骨细胞在支架内 的分布情况,并选择MTT试验来检测细胞活力。灌注组的SEM图片和组织学切片显示出更好的三维细胞生长以及支架内广泛的细胞分布;而在对照组中,软骨细胞仅在支架表面分散地形成单层。相应地,灌注组的MTT结果远高于对照组(0.123对0.067,P<0.01)。PEOT/PBT支架内的连续介质灌注有效地结合了软骨细胞接种和培养系统,用于组织工程气管的重建。

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