Ding Vanessa, Choo Andre B H, Oh Steve K W
Stem Cell Group, Bioprocessing Technology Institute, 20 Biopolis Way, #06-01 Centros, 138668, Singapore, Singapore.
Biotechnol Lett. 2006 Apr;28(7):491-5. doi: 10.1007/s10529-006-0005-8.
Development of a serum free, feeder-free (SFFF) culture platform for human embryonic stem cells (hESC) will be important for the expansion of hESC for future cell therapy applications. However, currently, culture of hESC consists of a combination of basal media, basic fibroblast growth factor (bFGF), serum replacer (SR) and conditioned media (CM) from feeders, and it is unclear which components of the mixture are absolutely critical in the maintenance of hESC. To evaluate the relative contributions of these media components in the development of SFFF culture, each was systematically eliminated and pluripotency assayed by dual embryonic stem cell markers, Oct-4 and TRA-1-60. We concluded that SR was the most critical component in the platform, followed by bFGF and CM produced by feeders, where down-regulation of Oct-4 occurred after 2, 5 and 5 passages, respectively, upon their withdrawal from the complete media.
开发一种用于人类胚胎干细胞（hESC）的无血清、无饲养层（SFFF）培养平台，对于未来细胞治疗应用中hESC的扩增至关重要。然而，目前hESC的培养由基础培养基、碱性成纤维细胞生长因子（bFGF）、血清替代物（SR）和来自饲养层的条件培养基（CM）组成，尚不清楚该混合物中的哪些成分对于hESC的维持绝对关键。为了评估这些培养基成分在SFFF培养发展中的相对贡献，我们对每种成分进行了系统去除，并通过双胚胎干细胞标志物Oct-4和TRA-1-60检测多能性。我们得出结论，SR是该平台中最关键的成分，其次是bFGF和饲养层产生的CM，当从完全培养基中分别去除它们后，Oct-4在第2、5和5代后分别出现下调。
