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Considerations on the use of ear chondrocytes as donor chondrocytes for cartilage tissue engineering

机译:关于将软骨细胞用作软骨组织工程的供体软骨细胞的考虑

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Articular cartilage is often used for research on cartilage tissue engineering. However, ear cartilage is easier to harvest, with less donor-site morbidity. The aim of this study was to evaluate whether adult human ear chondrocytes were capable of producing cartilage after expansion in monolayer culture. Cell yield per gram of cartilage was twice as high for ear than for articular cartilage. Moreover, ear chondrocytes proliferated faster. Cell proliferation could be further stimulated by the use of serum-free medium with Fibroblast Growth Factor 2 (FGF2) in stead of medium with 10% serum. To evaluate chondrogenic capacity, multiplied chondrocytes were suspended in alginate and implanted subcutaneously in athymic mice. After 8 weeks the constructs demonstrated a proteoglycan-rich matrix that contained collagen type Ⅱ. Constructs of ear chondrocytes showed a faint staining for elastin. Quantitative RT-PCR revealed that expression of collagen type Ⅱ was 2-fold upregulated whereas expression of collagen type Ⅰ was 2-fold down regulated in ear chondrocytes expanded in serum-free medium with FGF2 compared to serum-containing medium. Expression of alkaline phosphatase and collagen type X were low indicating the absence of terminal differentiation. We conclude that ear chondrocytes can be used as donor chondrocytes for cartilage tissue engineering. Furthermore, it may proof to be a promising alternative cell source to engineer cartilage for articular repair.
机译:关节软骨通常用于软骨组织工程研究。但是,耳软骨更容易收获,且供体部位的发病率更低。这项研究的目的是评估成年人耳软骨细胞在单层培养中扩增后是否能够产生软骨。耳朵每克软骨的细胞产量是关节软骨的两倍。此外,耳软骨细胞增殖更快。通过使用含成纤维细胞生长因子2(FGF2)的无血清培养基代替含10%血清的培养基,可以进一步刺激细胞增殖。为了评估软骨形成能力,将成倍的软骨细胞悬浮在藻酸盐中,皮下植入无胸腺小鼠中。 8周后,构建体显示出富含蛋白聚糖的基质,其中包含Ⅱ型胶原。耳软骨细胞的构建体显示出弹性蛋白的微弱染色。定量RT-PCR显示,与不含血清的培养基相比,在无血清的FGF2培养基中扩增的耳软骨细胞中,Ⅱ型胶原的表达上调了2倍,而Ⅰ型胶原的表达下​​调了2倍。碱性磷酸酶和X型胶原的表达较低,表明没有终末分化。我们得出结论,耳软骨细胞可用作软骨组织工程的供体软骨细胞。此外,它可能被证明是工程化软骨以进行关节修复的有前途的替代细胞来源。

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