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用于软骨组织工程的猪耳软骨祖细胞的分离与软骨分化。

Isolation and chondrogenic differentiation of porcine perichondrial progenitor cells for the purpose of cartilage tissue engineering.

机构信息

Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany.

出版信息

Cells Tissues Organs. 2013;198(3):179-89. doi: 10.1159/000354897. Epub 2013 Oct 18.

Abstract

In vivo, cartilage has a limited regenerative capacity. Clinical replacement strategies require a suitable cell source to provide a stable chondrocyte phenotype without hypertrophic cartilage development, while being broadly available, and harboring a high proliferative potential. Thus, the aim of this study was to analyze the proliferation and chondrogenic differentiation capacity of porcine perichondrial progenitor cells (PPC) isolated from auricular (ePPC) and tracheal cartilage (tPPC) as an alternative cell source to mesenchymal stem cells (MSC). The proliferative potential of these cell types was analyzed by means of doubling times. Cell pellets were cultured in chondrogenic differentiation medium for 4 weeks. Potential chondrogenic differentiation was investigated by histology and immunohistology in addition to gene expression analysis of the cartilage markers collagen II, aggrecan, cartilage oligomeric matrix protein (COMP), the precartilage marker collagen I, and the hypertrophic cartilage marker collagen X. PPC showed a proliferative behavior comparable to that of MSC. Chondrogenic stimulation resulted in a higher expression of collagen II, aggrecan, and COMP in ePPC as compared to tPPC and MSC, whereas the expression of collagen I was comparable in all cell types independently of differentiation stimulation. Collagen type X, however, could not be detected. The production of cartilage-like extracellular matrix components in PPC pellets was confirmed by histological and immunohistological stains. Elastin, a component of auricular cartilage, however, was not detected in ePPC-derived pellets. Thus, PPC present a promising cell source for tissue engineering of cartilage. Furthermore, ePPC may be more convenient than tPPC due to their higher chondrogenic potential and better accessibility.

摘要

在体内,软骨的再生能力有限。临床替代策略需要合适的细胞来源,以提供稳定的软骨细胞表型,而不会发生肥大性软骨发育,同时广泛可用,并具有高增殖潜力。因此,本研究旨在分析从耳软骨(ePPC)和气管软骨(tPPC)分离的猪软骨膜祖细胞(PPC)的增殖和软骨分化能力,作为间充质干细胞(MSC)的替代细胞来源。通过倍增时间分析这些细胞类型的增殖潜力。细胞团块在软骨分化培养基中培养 4 周。通过组织学和免疫组织化学以及软骨标志物胶原 II、聚集蛋白聚糖、软骨寡聚基质蛋白(COMP)、前软骨标志物胶原 I 和肥大性软骨标志物胶原 X 的基因表达分析来研究潜在的软骨分化。PPC 的增殖行为与 MSC 相当。软骨刺激导致 ePPC 中胶原 II、聚集蛋白聚糖和 COMP 的表达高于 tPPC 和 MSC,而胶原 I 的表达在所有细胞类型中均相似,与分化刺激无关。然而,不能检测到胶原 X。PPC 小球中软骨样细胞外基质成分的产生通过组织学和免疫组织化学染色得到证实。然而,在 ePPC 衍生的小球中未检测到耳软骨的弹性蛋白等成分。因此,PPC 为软骨组织工程提供了一种有前途的细胞来源。此外,由于 ePPC 具有更高的软骨形成潜力和更好的可及性,因此可能比 tPPC 更方便。

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