National Laboratory for Embryonic Stem Cell Research, Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
Tissue Eng Part C Methods. 2013 Feb;19(2):146-55. doi: 10.1089/ten.TEC.2012.0141. Epub 2012 Oct 16.
Currently, stem cell research faces a major bottleneck related to the low efficiency of methods to produce large quantities of human embryonic stem cells (ESC) for use in clinical trials. Most culture media currently employed for human ESC cultivation contain animal compounds, and cells are grown in static flasks. Besides the immediate contamination with nonhuman compounds, cell expansion in flasks tends to be laborious and nonefficient. Here we cultured human ESC in stirred microcarrier (MC) systems using an animal/human-component-free medium, to overcome both issues. The method developed to culture cells on suspended beads combined the use of polymeric MCs in stirred vessels with an optimized culture medium free of supplements of animal and human origin. This approach generated approximately 160 million cells within 6 days, which were shown to remain pluripotent. The process developed herein provides a step forward toward therapy due to the economic advantages in the production of human ESC and to their consequent low immunogenic potential.
目前，干细胞研究面临一个主要的瓶颈，即如何高效地生产大量用于临床试验的人类胚胎干细胞（ESC）。目前用于人类 ESC 培养的大多数培养基都含有动物成分，细胞在静态培养瓶中生长。除了立即受到非人类化合物的污染外，细胞在培养瓶中的扩增往往既费力又低效。在这里，我们使用无动物/人成分的培养基在搅拌微载体（MC）系统中培养人类 ESC，以克服这两个问题。在悬浮珠粒上培养细胞的方法结合了搅拌容器中使用聚合物 MC 和不含动物和人源补充剂的优化培养基。该方法在 6 天内产生了大约 1.6 亿个细胞，这些细胞显示出多能性。由于生产人类 ESC 的经济优势及其随之而来的低免疫原性潜力，本文所开发的方法为治疗提供了一个前进的步骤。
