In vitro redifferentiation of culture-expanded rabbit and human auricular chondrocytes for cartilage reconstruction.
In vitro redifferentiation of culture-expanded rabbit and human auricular chondrocytes for cartilage reconstruction.
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DOI：
10.1097/00006534-200102000-00020
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发表时间：
2001-02
影响因子：
3.6
通讯作者：
G. V. Osch;Simone W. van der Veen;H. L. Verwoerd-Verhoef
中科院分区：
文献类型：
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作者：
G. V. Osch;Simone W. van der Veen;H. L. Verwoerd-Verhoef
为了利用组织工程构建自体软骨移植物，细胞必须在体外增殖；然后它们就失去了软骨特有的表型。本研究的目的是评估增殖的耳软骨细胞使用各种培养条件重新表达其软骨表型的能力。从三只幼兔和三只成年兔子的耳朵软骨中分离出细胞，在单层培养中增殖后，将它们接种到藻酸盐中，并在含有三种不同剂量组合的胰岛素样生长因子 1 (IGF-1) 和转化生长因子-β2 (TGF-β2) 的无血清培养基中培养 3 周。作为对照，细胞在10%胎牛血清中培养，之前的实验证明其无法诱导再分化。当血清被胰岛素样生长因子-1 和转化生长因子-β2 取代时，来自幼年兔子（而非成年兔子）耳朵的软骨细胞合成了显着更多的糖胺聚糖。 II 型胶原蛋白阳性细胞的数量在年轻细胞中从 10% 增加到 97%，在成年细胞中增加到 33%。使用来自 12 名患者（年龄 7 至 60 岁）的人耳细胞，通过用胰岛素样生长因子和转化生长因子-β 代替血清也可以刺激糖胺聚糖的合成。尽管在这些条件下II型胶原蛋白阳性细胞的数量可以增加，但从未达到10%以上。来自五名患者的数据表明，通过添加 ITS+ 和皮质醇进一步优化培养条件，糖胺聚糖合成和 II 型胶原表达显着增加（两倍或三倍）。总之，本研究展示了一种在增殖的耳软骨细胞中恢复软骨表型的方法。这提高了生成用于重建外耳或修复鼻中隔缺损的人类软骨移植物的机会。
To construct an autologous cartilage graft using tissue engineering, cells must be multiplied in vitro; they then lose their cartilage-specific phenotype. The objective of this study was to assess the capacity of multiplied ear chondrocytes to re-express their cartilage phenotype using various culture conditions. Cells were isolated from the cartilage of the ears of three young and three adult rabbits and, after multiplication in monolayer culture, they were seeded in alginate and cultured for 3 weeks in serum-free medium with insulin-like growth factor 1 (IGF-1) and transforming growth factor-beta2 (TGF-beta2) in three different dose combinations. As a control, cells were cultured in 10% fetal calf serum, which was demonstrated in previous experiments to be unable to induce redifferentiation. Chondrocytes from the ears of young, but not adult, rabbits, synthesized significantly more glycosaminoglycan when serum was replaced by insulin-like growth factor-1 and transforming growth factor-beta2. The number of collagen type II-positive cells was increased from 10 percent to 97 percent in young cells and to 33 percent in adult cells. Using human ear cells from 12 patients (aged 7 to 60 years), glycosaminoglycan synthesis could also be stimulated by replacing serum with insulin-like growth factor and transforming growth factor-beta. Although the number of collagen type II-positive cells could be increased under these conditions, it never reached above 10 percent. Data from five patients showed that further optimization of the culture conditions by adding ITS+ and cortisol significantly increased (doubled or tripled) both glycosaminoglycan synthesis and collagen type II expression. In conclusion, this study demonstrates a method to regain cartilage phenotype in multiplied ear cartilage cells. This improves the chances of generating human cartilage grafts for the reconstruction of external ears or the repair of defects of the nasal septum.