Abstract
Background aims. Human embryonic stem (hES) cells hold great potential for cell therapy and regenerative medicine because of their pluripotency and capacity for self-renewal. The conditions used to derive and culture hES cells vary between and within laboratories depending on the desired use of the cells. Until recently, stem cell culture has been carried out using feeder cells, and culture media, that contain animal products. Recent advances in technology have opened up the possibility of both xeno-free and feeder-free culture of stem cells, essential conditions for the use of stem cells for clinical purposes. To date, however, there has been limited success in achieving this aim. Methods, results and conclusions. Protocols were developed for the successful derivation of two normal and three specific mutation-carrying (SMC) (Huntington's disease and myotonic dystrophy 1) genomically stable hES cell lines, and their adaptation to feeder-free culture, all under xeno-free conditions.
Acknowledgments
This work was supported by Medical Research Council UK grants.
Author contributions: DI, ES and PB designed the methods; ES, VW, JL, DS, AP, NK, SC and MS performed experiments; GC obtained patient consents; HP and CO were responsible for genomic analyzes; ES, HP and MS contributed to the writing of the methods; DI, CO and PB wrote the manuscript.
Competing financial interests: The authors have declared that no competing interests exist.