Objective To find a new source to produce hESC lines. Methods D3 embryos with low morphological scores were cultured to blastocyst stage. Trophectoderm cells were separated from the ICMs by immunosurgery and isolated ICMs were cultured for 58 days on mitomycintreated mouse embryonic fibroblasts (MEFs). Colonies derived from the ICMs were passed every 47 days and evaluated for cell surface markers including AKP, OCT4, SSEA4, SSEA1, TRA160, TRA181, differentiation potentials and karyotypes. Results A total of 19 blastocysts were obtained from 130 embryos (quality score < 16) and blastulation rate was 1462%. Immunosurgery resulted in the formation of 5 primary colonies, which produced 2 cell lines. Both cell lines satisfied the criteria that characterize pluripotent hESC. Conclusion A subset with poor quality D3 embryos judged on the basis of morphological assessment