With their counterparts at comparable stages of development in vivo and putative paracrine factors secreted by the reproductive tract were shown to enhance early embryonic development. Recent progress in sequential culture media allowed extended culture of human embryos to the blastocyst-stage and blastocyst transfer is effective in selecting high-quality embryos for successful pregnancy, leading to the birth of several million IVF babies. However, the efficiency of human blastocyst development in vitro remains to be improved. Several studies using surplus human material suggested the promotion of blastocyst development in vitro when culture media were supplemented with growth factors, including EGF, IGF-I, BDNF, and granulocyte macrophage colony-stimulating factor. Because most routine human embryo cultures do not contain growth factors, we hypothesized that inclusion of autocrine/paracrine growth factors in the culture media could improve early embryonic development. We investigated the expression of key Crizotinib ligand-receptor pairs in cleavage-stage human embryos derived from tri-pronuclear zygotes and specific ligands in human uterine endometrium. We individually cultured abnormally fertilized zygotes and normally fertilized day 3 embryos from IVF programs in micro-drops using routine serum-free culture media supplemented with a group of key growth factors to improve embryo development. We examined if these growth factors could improve blastocyst outgrowth in vitro using normally fertilized hatching blastocysts. Because the development of reconstructed embryos after SCNT is sub-optimal, we also tested if supplementation of culture media with key growth factors could increase the development of SCNT-derived embryos. Following culturing individual human embryos in chemically defined serum-free media, we demonstrated the ability of key autocrine/paracrine growth factors to promote early embryonic development and implantation. Treatment with key growth factors enhanced the development of abnormal tri-pronuclear zygotes, normally fertilized human embryos, and reconstructed embryos following SCNT. The key growth factors not only stimulated embryo growth but also increased the proportion of morphologically good blastocysts, suggesting improvement of embryo quality. Although the facilitatory effects of individual growth factors on human embryo development have been reported, the present use of multiple growth factors likely exerts overlapping and redundant actions to allow optimal early embryo development. Culturing of individual embryos further minimizes cross-interaction of embryos and provides the basis to monitor the functions of single high quality embryos for transfer. Demonstration of the facilitatory effects of key growth factors to promote blastocyst outgrowth further provides future opportunity to include them in embryo transfer media to improve implantation success. Our immunofluorescence staining and real-time RT-qPCR analyses confirmed the expression of key ligand-receptor pairs in human early embryos, underscoring their importance as autocrine/paracrine factors. For each ligand-receptor pairs, ligand antigens were found in the cytoplasm of blastomeres whereas the receptor antigens were found in the plasma membrane, suggesting the secretion of these paracrine/autocrine ligands to act on membrane receptors in an autocrine/paracrine manner. As a 3- day-old human embryo enters the uterus at the morula stage, further development could also be regulated by paracrine factors secreted by the endometrium. Our RT-PCR and immunostaining studies confirmed the expression of these key growth factors in the human endometrium, suggesting their paracrine roles in support of embryo growth after the morula stage.