title: Promotion of human early embryonic development and blastocyst outgrowth in vitro using autocrine/paracrine growth factors.
abstract: Studies using animal models demonstrated the importance of autocrine/paracrine factors secreted by preimplantation embryos and reproductive tracts for embryonic development and implantation. Although in vitro fertilization-embryo transfer (IVF-ET) is an established procedure, there is no evidence that present culture conditions are optimal for human early embryonic development. In this study, key polypeptide ligands known to be important for early embryonic development in animal models were tested for their ability to improve human early embryo development and blastocyst outgrowth in vitro. We confirmed the expression of key ligand/receptor pairs in cleavage embryos derived from discarded human tri-pronuclear zygotes and in human endometrium. Combined treatment with key embryonic growth factors (brain-derived neurotrophic factor, colony-stimulating factor, epidermal growth factor, granulocyte macrophage colony-stimulating factor, insulin-like growth factor-1, glial cell-line derived neurotrophic factor, and artemin) in serum-free media promoted >2.5-fold the development of tri-pronuclear zygotes to blastocysts. For normally fertilized embryos, day 3 surplus embryos cultured individually with the key growth factors showed >3-fold increases in the development of 6-8 cell stage embryos to blastocysts and >7-fold increase in the proportion of high quality blastocysts based on Gardner's criteria. Growth factor treatment also led to a 2-fold promotion of blastocyst outgrowth in vitro when day 7 surplus hatching blastocysts were used. When failed-to-be-fertilized oocytes were used to perform somatic cell nuclear transfer (SCNT) using fibroblasts as donor karyoplasts, inclusion of growth factors increased the progression of reconstructed SCNT embryos to >4-cell stage embryos. Growth factor supplementation of serum-free cultures could promote optimal early embryonic development and implantation in IVF-ET and SCNT procedures. This approach is valuable for infertility treatment and future derivation of patient-specific embryonic stem cells.
authors.affiliation: Program of Reproductive and Stem Cell Biology, Department of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Kawamura
authors.forename: Yuan
authors.initials: Y
authors.lastname: Chen
authors.forename: Yimin
authors.initials: Y
authors.lastname: Shu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Cheng
authors.forename: Jie
authors.initials: J
authors.lastname: Qiao
authors.forename: Barry
authors.initials: B
authors.lastname: Behr
authors.forename: Renee A Reijo
authors.initials: RA
authors.lastname: Pera
authors.forename: Aaron J W
authors.initials: AJ
authors.lastname: Hsueh
mesh_terms: D000328:Adult; D019898:Autocrine Communication / Q000187:drug effects*; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015925:Cryopreservation; D003470:Culture Media / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects*; D004717:Endometrium / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D008024:Ligands; D053652:Nuclear Transfer Techniques; D019899:Paracrine Communication / Q000187:drug effects*; D011956:Receptors, Cell Surface / Q000378:metabolism; D015053:Zygote / Q000166:cytology / Q000187:drug effects / Q000254:growth & development
chemical_list: D003470:Culture Media; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D011956:Receptors, Cell Surface