Derivation and Expansion of Human Embryonic Stem Cells Under Xeno-Free, Defined Conditions
Human embryonic stem cells (hESCs) hold great promise in regenerative medicine and cell therapy due to their unique properties: unlimited self-renewal and the pluripotency to differentiate into all cell lineages in the body. However, the overwhelming majority of currently available hESC lines have been directly or indirectly exposed to materials containing animal-derived components during their derivation, propagation, and cryopreservation. The use of animal-derived components would prevent the use of hESCs for clinical purposes, due to the possibility of xenogeneic bimolecule and pathogen contamination. Therefore, the establishment of clinical-grade hESC lines in xeno-free, chemically defined conditions is the first and key step. In this chapter, we review and summarize the history and current state of derivation, propagation and expansion of hESCs in static and suspension cultures in xeno-free, defined conditions. The main part of this review focuses on the recent advances in the generation and expansion of hESCs in xeno-free, chemically defined conditions. Based on previous studies, we also put forward the possible means for deriving and expanding hESC lines in xeno-free, defined conditions under current good manufacturing process (cGMP) standards that will enable the generation of clinical-grade hESC lines for the clinical purposes.
KeywordsClinical-Grade hESCs Culture and expansion Dissociation Human embryonic stem cells (hESCs) Passaging Suspension culture Xeno-Free (XF)
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