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表面抗原是否适合用于验证基于细胞的植入物中人类软骨细胞的再分化潜能和培养纯度?

Are surface antigens suited to verify the redifferentiation potential and culture purity of human chondrocytes in cell-based implants.

作者信息

Krüger M, Krüger J P, Kinne R W, Kaps C, Endres M

机构信息

TransTissue Technologies GmbH, Berlin, Germany.

TransTissue Technologies GmbH, Berlin, Germany; Department of Rheumatology, Laboratory for Tissue Engineering, Charité - Universitätsmedizin Berlin, Berlin, Germany.

出版信息

Tissue Cell. 2015 Oct;47(5):489-97. doi: 10.1016/j.tice.2015.07.004. Epub 2015 Jul 17.

Abstract

Cell expansion in vitro is a prequisite to obtain a sufficient quantity of cells for cell-based cartilage repair of articular cartilage lesions. During this process verification of redifferentiation potential of highly expanded chondrocytes is required. Furthermore, cellular impurities of chondrocyte cultures have to be excluded. For this purpose, redifferentiation of expanded human chondrocytes in passage 3 or 5 was initiated in bioresorbable polyglycolic acid-fibrin (PGA-fibrin) scaffolds and selected potential markers were analysed during the process of cell expansion and redifferentiation. Chondrocyte expansion was accompanied by a decrease of collagen type II and COMP and an increase of collagen type I expression indicating cell dedifferentiation. Redifferentiation of chondrocytes in PGA-fibrin scaffolds was accompanied by an increase of collagen II/I ratio. Flow cytometric analyses revealed that in contrast to CD44 and CD49e, CD63 and CD166 showed significant changes in the number of positive cells during redifferentiation. CD14 and CD45 are not expressed by chondrocytes and are therefore possible candidates to detect specifically monocytes or haematopoetic cells in chondrocyte cultures. Characterization of surface antigen expression revealed two promising candidates (CD63 and CD166) to describe the process of redifferentiation, while CD14 and CD45 are suitable markers to exclude impurities by monocytes or haematopoetic cells.

摘要

体外细胞扩增是为关节软骨损伤的基于细胞的软骨修复获得足够数量细胞的前提条件。在此过程中,需要验证高度扩增的软骨细胞的再分化潜能。此外,必须排除软骨细胞培养物中的细胞杂质。为此,在可生物降解的聚乙醇酸 - 纤维蛋白(PGA - 纤维蛋白)支架中启动第3代或第5代扩增的人软骨细胞的再分化,并在细胞扩增和再分化过程中分析选定的潜在标志物。软骨细胞扩增伴随着II型胶原蛋白和软骨寡聚基质蛋白的减少以及I型胶原蛋白表达的增加,表明细胞去分化。PGA - 纤维蛋白支架中软骨细胞的再分化伴随着II/I型胶原蛋白比例的增加。流式细胞术分析显示,与CD44和CD49e相比,CD63和CD166在再分化过程中阳性细胞数量有显著变化。软骨细胞不表达CD14和CD45,因此它们可能是检测软骨细胞培养物中特异性单核细胞或造血细胞的候选物。表面抗原表达的表征揭示了两个有前景的候选物(CD63和CD166)来描述再分化过程,而CD14和CD45是排除单核细胞或造血细胞杂质的合适标志物。

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