Challenges and approaches to the culture of pluripotent human embryonic stem cells.

Since the establishment of the first human embryonic stem cell (hESC) lines, several groups have described the derivation and culture of hESC lines in various culture conditions. In this review, we describe how hESC lines have been derived from the inner cell mass of blastocysts or morula-stage embryos and the culture conditions used. In order to be used for therapeutic purposes, the pluripotent hESC lines must be established and propagated according to good manufacturing practice quality requirements. In addition, any use of animal-derived components should be avoided to gain safer hESC lines for clinical purposes. Here, we will describe the development in derivation and chemically defined culturing conditions of hESC towards good manufacturing practice and discuss the future challenges for hESCs in clinical use. Similarly, we discuss the challenges and future directions in optimization of standard culture conditions of hESCs for research purposes.