Subcloning and Alternative Methods for the Derivation and Culture of Human Embryonic Stem Cells

Abstract

Human embryonic stem (ES) cell lines may have broad applications, including the study of development and the differentiation process, lineage commitment, self-maintenance, and precursor cell maturation. They may also serve as models in research done on the functions of genes and proteins, drug testing, and drug toxicity. The first human ES cells were derived by Thomson and colleagues (1) from the inner cell mass (ICM) of surplus blastocysts donated by couples undergoing in vitro fertilization treatments. These lines met most of the criteria for ES cell lines listed in Table 1, but their clonality was not tested in that study. Also, the ability of human ES cells to contribute to embryonic development in chimeric embryos cannot be examined for obvious ethical reasons. Since the first report on human ES cell derivation, several other groups have reported the derivation of additional lines (2–4) At present, there are more than 70 human ES cell lines in several laboratories around the world, according to a list published by the National Institutes of Health (NIH; http://www.nih.gov/news/stemcell/index/news/stemcell/index). Although the NIH list does not offer full information on all the lines fulfilling all the ES cell criteria listed in Table 1, it suggests that the derivation of human ES cells is a reproducible procedure with reasonable success rates.