Rapid and large-scale formation of chondrocyte aggregates by rotational culture
Rapid and large-scale formation of chondrocyte aggregates by rotational culture
复制标题
DOI：
10.3727/000000003108747037
复制
发表时间：
2003-01-01
影响因子：
3.3
通讯作者：
Tateishi, T
中科院分区：
文献类型：
--
作者：
Furukawa, KS;Suenaga, H;Tateishi, T
关节软骨中的软骨细胞合成 H 型胶原蛋白和大的硫酸化蛋白聚糖，而单层 (2D) 培养的相同细胞则去分化为成纤维细胞并表达 I 型胶原蛋白和小蛋白聚糖。另一方面，开发了颗粒培养系统作为防止软骨细胞表型调节和促进去分化细胞再分化的方法。由于颗粒培养系统通过离心机在每个管中仅形成一个细胞聚集体，因此颗粒不能用于生产组织工程软骨。因此，我们尝试通过旋转培养形成软骨细胞聚集体，期望一次性形成大量聚集体。为了增加细胞-细胞相互作用并减少软骨细胞-材料相互作用，使用蛋白质吸附保留和细胞粘附性低的培养皿。此外，尝试旋转摇动含有细胞的培养皿以增加细胞与细胞的相互作用。振荡速度设置为 80 rpm，因此细胞将分布在培养皿的中心，以增加细胞与细胞接触的频率。在这些条件下，牛关节软骨细胞在几个小时内开始聚集。在24-36It的旋转培养下，观察到具有光滑表面的聚集体。参数例如培养时间的增加和TGF-β的添加控制聚集体的直径。聚集体的外围有许多梭形细胞，细胞在横截面上倾向于形成多层区。此外，还观察到与颗粒培养物几乎相同的腔隙状结构。结果发现，聚集体的内部结构与之前报道的颗粒的内部结构相似。因此，旋转培养形成的聚集体可能成为制造组织工程人工软骨的重要组成部分。
Chondrocytes in articular cartilage synthesize collagen type H and large sulfated proteoglycans, whereas the same cells cultured in monolayer (2D) dedifferentiate into fibroblastic cells and express collagen type I and small proteoglycans. On the other hand, a pellet culture system was developed as a method for preventing the phenotypic modulation of chondrocytes and promoting the redifferentiation of dedifferentiated ones. Because the pellet culture system forms only one cell aggregate each tube by a centrifugator, the pellet could not be applied to produce a tissue-engineered cartilage. Therefore, we tried to form chondrocyte aggregates by a rotational culture, expecting to form a large number of aggregates at once. In order to increase cell-cell interactions and decrease chondrocyte-material interaction, dishes with low retention of protein adsorption and cell adhesiveness were used. In addition, rotational shaking of the dish including cells was attempted to increase the cell-cell interaction. The shaking speed was set at 80 rpm, so the cells would be distributed in the center of the dish to augment the frequency of cell-cell contact. Under these conditions, bovine articular chondrocytes started aggregating in a few hours. At 24-36 It of rotational culture, aggregates with smooth surfaces were observed. Parameters such as increase of culture time and addition of TGF-beta controlled diameters of the aggregates. There were many fusiform cells at the periphery of the aggregates, where the cells tended to form a multilayered zone in cross sections. In addition, lacune-like structure, which was almost the same as pellet culture, was observed. It was found that the internal structure of the aggregates was similar to that of pellets reported previously. Therefore, the aggregates formed by a rotational culture could become an essential component to make tissue-engineered artificial cartilage.