Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland, Australia.
J Orthop Res. 2011 Apr;29(4):539-46. doi: 10.1002/jor.21264. Epub 2010 Oct 18.
Expanding human chondrocytes in vitro while maintaining their ability to form cartilage remains a key challenge in cartilage tissue engineering. One promising approach to address this is to use microcarriers as substrates for chondrocyte expansion. While microcarriers have shown beneficial effects for expansion of animal and ectopic human chondrocytes, their utility has not been determined for freshly isolated adult human articular chondrocytes. Thus, we investigated the proliferation and subsequent chondrogenic differentiation of these clinically relevant cells on porous gelatin microcarriers and compared them to those expanded using traditional monolayers. Chondrocytes attached to microcarriers within 2 days and remained viable over 4 weeks of culture in spinner flasks. Cells on microcarriers exhibited a spread morphology and initially proliferated faster than cells in monolayer culture, however, with prolonged expansion they were less proliferative. Cells expanded for 1 month and enzymatically released from microcarriers formed cartilaginous tissue in micromass pellet cultures, which was similar to tissue formed by monolayer-expanded cells. Cells left attached to microcarriers did not exhibit chondrogenic capacity. Culture conditions, such as microcarrier material, oxygen tension, and mechanical stimulation require further investigation to facilitate the efficient expansion of clinically relevant human articular chondrocytes that maintain chondrogenic potential for cartilage regeneration applications. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 29:539-546, 2011.
在体外扩增人类软骨细胞的同时保持其形成软骨的能力仍然是软骨组织工程的一个关键挑战。一种有前途的方法是使用微载体作为软骨细胞扩增的基质。虽然微载体已显示出对动物和异位人软骨细胞扩增的有益效果，但它们对新鲜分离的成人关节软骨细胞的应用尚未确定。因此，我们研究了这些临床上相关的细胞在多孔明胶微载体上的增殖和随后的软骨分化，并将其与使用传统单层培养的细胞进行了比较。软骨细胞在 2 天内附着在微载体上，并在旋转瓶中培养 4 周以上仍保持存活。微载体上的细胞呈现出展开的形态，最初比单层培养中的细胞增殖更快，然而，随着时间的延长，它们的增殖能力下降。在微载体上扩增 1 个月并通过酶从微载体上释放的细胞在微团滴培养中形成软骨组织，这与单层扩增细胞形成的组织相似。留在微载体上的细胞没有表现出软骨生成能力。培养条件，如微载体材料、氧张力和机械刺激，需要进一步研究，以促进临床相关的人关节软骨细胞的有效扩增，这些细胞保持软骨生成潜力，用于软骨再生应用。©2010 矫形研究协会。由 Wiley Periodicals, Inc. 出版。J Orthop Res 29:539-546, 2011.
