Abstract: The various embodiments herein provide a method for derivation and long term establishment of ground state pluripotent embryonic stem cells. Further the embodiments herein provides a method to inhibit the ERK and TGF ? signalling pathways for long term maintenance of the embryonic stem cells. The R2i mouse embryonic stem (ES) cells are derived from 3.5 day blastocysts. The mouse ES cells are cultured in media containing R2i and 2i inhibitors of ERK and TGF ? pathways. The ES cells are subjected to in vitro and in vivo differentiation. The ES cells are subjected to RT-PCR and qRT-PCR, flow cytometry and karyotyping. The result reveals that the R2i maintains the ground state of ES cells and self renewal. Also R2i increases embryonic cleavage and clonal propagation of ES. Further R2i asserts genomic integrity and pluripotency of ES.