Implantation of blastocysts is a well-organized process regulated by multiple growth factors and cytokines. We demonstrated the facilitatory effects of key growth factors to promote blastocyst outgrowth. The trophectoderm cells of blastocysts differentiate during embryonic development to form the invasive trophoblasts that mediate implantation of embryos into the uterine wall. The outgrowth of trophoblast cells from cultured blastocysts is believed to reflect the proper differentiation of the embryo, important for trophoblast invasion of the endometrial stroma during implantation in utero. Although blastocyst transfer is effective to select the best quality embryos with high implantation potential, overall implantation rate is,30%, suggesting human embryo transfer might be improved. Due to the low amount of liquid in the uterine cavity, factors included in the transfer media could be retained in high concentrations. Indeed, embryo transfer in medium containing hyaluronan is effective in improving implantation rates in patients with recurrent implantation failure.Hyaluronan is the major glycosaminoglycan present in LDK378 follicular, oviductal and uterine fluids and presumably promotes embryo–endometrial interactions during the initial phases of implantation. Because key growth factors promoted blastocyst outgrowth in vitro, future supplementation of embryo transfer media with key growth factors could also promote implantation during embryo transfer. Generating an autologous patient-specific embryonic stem cell line from SCNT embryos holds great promise for the treatment of degenerative human diseases. Successful derivation of embryonic stem cell lines following SCNT has been reported in mouse, rabbit, and non-human primates. However, the efficiency for the production of embryonic stem cell lines following SCNT is still low, particularly when adult somatic cells were used as the donor karyoplasts. Although many embryonic stem cell lines have been derived from surplus human blastocysts, no human cell-lines have been generated following SCNT. Among the many compounding factors, suboptimal in vitro culture condition contributes to the poor embryonic development of reconstructed embryos following SCNT. The present study represents an attempt to optimize the culture conditions for the development of human SCNT embryos. Although no blastocyst was obtained following fibroblast nuclear transfer, there was a trend to an augmented development of reconstructed embryos cultured with media containing autocrine/paracrine growth factors. Results from the present study provide the basis for future use of autocrine/paracrine factors to facilitate the derivation of patient-specific embryonic stem cells. In conclusion, the present study demonstrated the utility of growth factor supplementation for optimal human early embryo development and blastocyst outgrowth. The findings may allow the design of better conditions for individual human embryo cultures, for estimating their developmental potentials using secretory products, and for the inclusion of growth factors in embryo transfer media to promote implantation. Although the present experimental design is based on the supplementation of endogenous growth factors diluted during assisted reproductive procedures, future studies on the potential side effects of these paracrine/autocrine factors on chromosomal numbers, genomic integrity and epigenetic modifications are essential before clinical use. The gut microbiota performs necessary metabolic functions such as production of short chain fatty acids and synthesis of vitamins. It also influences the maturation of the immune system after birth, which is clearly illustrated in studies of germ-free animals. GF mice have fewer intestinal dendritic cells and mice with a restricted microbiota have less plasmacytoid DCs.