Abstract：
Allogeneic serum and tissue-specific extracellular matrix have been shown to maintain permanently differentiated cell phenotype in culture. This is of particular importance for human tenocytes, a cell population that readily loses its function during ex vivo culture. With these in mind, herein we extracted human tenocytes using either foetal bovine serum or human serum, cultured them in the absence and presence of carrageenan and Ficoll®, the most widely used macromolecular crowding agents (to induce tissue-specific extracellular matrix deposition), and assessed cellular function, via metabolic activity, viability, proliferation and immunofluorescence for collagen related molecules, non-collagenous molecules and transmembrane molecules. At day 7, longest time point assessed, neither carrageenan nor Ficoll® significantly affected metabolic activity, viability and proliferation in either serum and human serum significantly increased metabolic activity and proliferation. At day 7, in the absence of macromolecular crowding, cells in human serum deposited significantly lower collagen type VI, biglycan, versican and tenomodulin than cells in foetal bovine serum. Interestingly, at day 7, in comparison to the no macromolecular crowding group, carrageenan in foetal bovine serum induced the highest effect, as judged by the highest number of significantly increased molecules (collagen type I, collagen type IV, collagen type V, collagen type VI, transforming growth factor β1, matrix metalloproteinase 14, lumican, versican, scleraxis and integrin α2β1). These data, although contradict previous observations where human serum outperformed foetal bovine serum, at the same time, support the use of foetal bovine serum in the development of cell-based medicines.
摘要：
已显示同种异体血清和组织特异性细胞外基质在培养物中保持永久分化的细胞表型。这对人类肌腱细胞特别重要，在离体培养过程中容易丧失功能的细胞群。考虑到这些，在本文中，我们使用胎牛血清或人血清提取人腱细胞，在不存在角叉菜胶和Ficoll®的情况下培养它们，最广泛使用的大分子拥挤剂（诱导组织特异性细胞外基质沉积），并评估细胞功能，通过代谢活动，生存能力，胶原蛋白相关分子的增殖和免疫荧光，非胶原分子和跨膜分子。在第7天，评估的最长时间点，角叉菜胶和Ficoll®都不显著影响代谢活动，血清和人血清中的活力和增殖显着增加了代谢活性和增殖。在第7天，在没有大分子拥挤的情况下，人血清中沉积的细胞显著降低了VI型胶原，biglycan,versican和腱调节蛋白比胎儿牛血清中的细胞。有趣的是,在第7天，与无大分子拥挤组相比，角叉菜胶对胎牛血清的诱导效果最高,根据最大数量的显著增加的分子(I型胶原蛋白，胶原蛋白IV型，胶原蛋白V型，胶原蛋白VI型，转化生长因子β1，基质金属蛋白酶14，Lumican，versican,巩膜和整合素α2β1)。这些数据,尽管与以前人类血清优于胎牛血清的观察结果相矛盾，同时,支持在细胞药物开发中使用胎牛血清。
