A Survey of Parameters Involved in the Establishment of New Lines of Human Embryonic Stem Cells
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Since the derivation of the first human embryonic stem cell (hESC) lines by Thomson and coworkers in 1998, more than 1,200 different hESC lines have been established worldwide. Nevertheless, there is still a recognized interest in the establishment of new lines of hESC, particularly from HLA types and ethnic groups currently underrepresented among the available lines. The methodology of hESC derivation has evolved significantly since 1998, when human LIF (hLIF) was used for maintenance of pluripotency. However, there are a number of different strategies for the several steps involved in establishing a new line of hESC. Here we make a survey of the most relevant parameters used between 1998 and 2010 for the derivation of the 375 hESC lines deposited in two international stem cell registries, and able to form teratomas in immunocompromised mice. Although we identify some trends in the methodology for establishing hESC lines, our data reveal a much greater heterogeneity of strategies than what is used for derivation of murine ESC lines, indicating that optimum conditions have not been consolidated yet, and thus, hESC establishment is still an evolving field of research.
KeywordsHuman embryonic stem cells Cell line derivation Pluripotency Human embryo Cell therapy
This work was supported by a grant from Conselho Nacional de Desenvolvimento Científico e Tecnológico/Departamento de Ciência e Tecnologia do Ministério da Saúde (CNPq/MS/DECIT). AMF, RS and ESSA have/had Ph.D. fellowships from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP). The authors declare no conflicts of interest.
- 21.Andrews, P. W., Stacey, G. N., & Sato, J. D. (editors) (2010). Special issue on novel human embryonic stem cell lines and related resources. In Vitro Cellular & Developmental Biology. Animal, 46(3–4), 167–402.Google Scholar
- 24.Fraga, A. M., Sukoyan, M., Rajan, P., et al. (2010). Establishment of a Brazilian line of human embryonic stem cells in defined medium—implications for cell therapy in an ethnically diverse population. Cell Transplantation, doi: 10.3727/096368910X522261.
- 35.Khoo, M. L., McQuade, L. R., Smith, M. S., Lees, J. G., Sidhu, K. S., & Tuch, B. E. (2005). Growth and differentiation of embryoid bodies derived from human embryonic stem cells: effect of glucose and basic fibroblast growth factor. Biology of Reproduction, 73(6), 1147–1156.PubMedCrossRefGoogle Scholar