The present disclosure provides novel culture system and methods for culturing and propagating hESCs in a substantially undifferentiated state for several passages. The ability to grow such cells without differentiation has important applications for therapeutic uses of ES cells for treating human disorders using tissue transplantation and/or gene therapy techniques. In particular, the disclosure further relates to conditioned medium obtained from human germ lineage derived feeder cells (GLDF). The hESC lines are derived, cultured and propagated in substantially undifferentiated state using the conditioned media from GLDF cells of the disclosure. In particular, the disclosure relates to the xeno-free derivation, culture and propagation of hESCs using conditioned medium of GLDF cells obtained thereof.
