Seeding Density Modulates Migration and Morphology of Rabbit Chondrocytes Cultured in Collagen Gels
Seeding Density Modulates Migration and Morphology of Rabbit Chondrocytes Cultured in Collagen Gels
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DOI：
10.1002/bit.22029
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发表时间：
2009-01-01
影响因子：
3.8
通讯作者：
Taya, Masahito
中科院分区：
文献类型：
--
作者：
Khoshfetrat, Ali Baradar;Kino-oka, Masahiro;Taya, Masahito
采用胶原凝胶包埋兔软骨细胞的培养方法，研究早期培养阶段的细胞行为和细胞聚集的结构。在1.0 X 10(5)个细胞/cm(3)时，植入的软骨细胞向纺锤形形态转变，并通过可能的迁移和聚集形成星形的松散聚集体。这些聚集体伴随着II型胶原蛋白的缺乏，而1.6 x 10(6)个细胞/cm 3的细胞表现出活跃的增殖，形成富含II型胶原蛋白的致密聚集体。在第5天进行立体观察，根据凝胶中单个细胞的球形度的形态学相关参数来定义迁移细胞。随着播种密度的增加，迁移细胞的频率呈下降趋势，而分裂细胞的频率呈相反趋势。1.0 × 10(5)个细胞/cm(3)时，细胞迁移频率为0.25，是1.6 × 10(6)个细胞/cm(3)时的25倍。此外，mRNA表达分析显示，与1.6 x 106细胞/cm的细胞相比，1.0 x 10(5)个细胞/cm的软骨细胞中与分化相关的II型胶原蛋白明显下调，与迁移相关的基质金属蛋白酶明显上调(3)。这些数据支持了不同种子密度培养中细胞迁移和聚集体形成的形态学分析。由此可见，种子密度是影响细胞行为和聚集体结构从而调节培养软骨质量的重要因素。Biotechnol。Bioeng。2009;102: 294 - 302。(C) 2008 Wiley期刊有限公司
The cultures of rabbit chondrocytes embedded in collagen gels were conducted to investigate the cell behaviors and consequent architectures of cell aggregation in an early culture phase. The chondrocyte cells seeded at 1.0 X 10(5) cells/cm(3) underwent a transition to spindle-shaped morphology, and formed the loose aggregates with a starburst shape by means of possible migration and gathering. These aggregates accompanied the poor production Of collagen type II, while the cells seeded at 1.6 x 10(6) cells/cm 3 exhibited active proliferation to form the dense aggregates rich in collagen type II. Stereoscopic observation was performed at 5 days to define the migrating cells in terms of a morphology-relating parameter of sphericity determined for individual cells in the gels. The frequency of migrating cells decreased with increasing seeding density, while the frequency of dividing cells showed the counter trend. The culture seeded at 1.0 x 10(5) Cells/cm(3) gave the migrating cell frequency of 0.25, the value of which was 25 times higher than that at 1.6 x 10(6) cells/cm(3). In addition, the analysis of mRNA expression revealed that the chondrocyte cells seeded at 1.0 x 10(5) cells/cm(3) showed appreciable down-regulation in collagen type II relating to differentiation and up-regulation in matrix metalloproteinases relating to migration, as compared to the cells seeded at 1.6 x 106 cells/cm(3). These data supports the morphological analyses concerning the cell migration and aggregate formation in the cultures with varied seeding densities. it is concluded that the seeding density is an important factor to affect the cell behaviors and architecture of aggregates and thereby to modulate the quality of cultured cartilage. Biotechnol. Bioeng. 2009;102: 294-302. (C) 2008 Wiley Periodicals, Inc.