海藻酸钠复合载体长期培养软骨细胞的生物学稳定性

背景：近年研究发现在海藻酸钠三维培养体系中软骨细胞存活时间延长,细胞外基质分泌旺盛。目的：观察海藻酸钠复合载体对体外长期培养软骨细胞生物学稳定性的影响。方法：将兔软骨细胞接种至以海藻酸钠、透明质酸、壳聚糖、纤维连接蛋白、碱性成纤维细胞生长因子为主要成分的复合载体三维培养体系中,以平面培养作为对照。定期观察在两种不同培养体系中软骨细胞形态学改变,细胞生长曲线差异,细胞上清液糖胺多糖含量差异。结果与结论：软骨细胞在海藻酸钠复合载体中生长良好,生长旺盛,并形成球状细胞团,细胞分裂增殖活跃;培养2d时复合载体培养软骨细胞增殖稍高于平面培养软骨细胞;培养4d时可见复合载体培养软骨细胞增殖明显加快;培养6~14d时复合载体培养的仍保持较稳定增殖,而平面培养的软骨细胞增殖逐渐降低;复合载体细胞外液糖胺多糖含量明显高于平面培养软骨细胞。说明海藻酸钠复合载体可以长期培养软骨细胞并保持其生物学稳定性。

Biological stability of long-term culture chondrocytes on three-dimensional culture system of multifactorial sodium alginate

BACKGROUND： Previous studies demonstrated that the survival time of chondrocytes is prolonged in three-dimensional culture system of multifactorial sodium alginate; extracellular matrix has a strong secretion. OBJECTIVE： To observe the effect of biological stability of long-term culture chondrocytes on three-dimensional culture system of multifactorial sodium alginate. METHODS： Chondrocytes were implanted on three-dimensional culture system which comprises sodium alginate, hyaluronic acid, chitosan, fibronectin, and basic fibroblast growth factor （bFGF）, and the plane culture as controls. The morphological changes, differences of cell growth curves, and the contents of glycosaminoglycan （GAG） of cells supernatant fluid of chondrocytes were regularly observed in two different culture systems. RESULTS AND CONCLUSION： Morphous of chondrocytes on three-dimensional culture system of multifactorial sodium alginate grew well and flourishingly, and formed the globular group of cells, cell proliferation was active. Proliferation of chondrocytes on the culture system of multifactorial sodium alginate was slightly higher than that implanted on plane cultured 2 days. Proliferation of chondrocytes was significantly higher cultured 4 days on the culture system of multifactorial sodium alginate. Chondrocytes were keep stable proliferation at 6-14 days culture; while the proliferation of chondrocytes cultured on plane was gradually decrease. The contents of GAG of cells supernatant fluid of the three-dimensional culture system were significantly higher than monolayer culture system. Chondrocytes on three-dimensional culture system of multifactorial sodium alginate can long-term culture and keep their biological stability.