title: The novel use of modified pig zygotic medium for the efficient culture of the preimplantation mouse embryos.
abstract: A high potassium concentration in culture media is considered detrimental to in vitro culture of mouse embryos. Here we show that pig zygotic medium (PZM) containing a higher concentration of potassium, and modified to contain 0.2 mM glucose and 0.01 mM EDTA, supported efficient pre- and post-implantation development of mouse zygotes to blastocysts and live pups, respectively. At first, modified PZM (mPZM) was compared with other culture media such as M16, CZB and KSOM-AA for its ability to support development of in vivo mouse zygotes to the blastocyst stage. The proportions of zygotes reaching 2-cell (94-99%) and blastocyst (90-96%) stages in mPZM and other media were not different. However, hatching rates of blastocysts were different (P < 0.05); whereas more than 90% of the blastocysts were hatching in mPZM or KSOM-AA, only 60% of the blastocysts did in M16 or CZB media (P < 0.05). Next we compared post-implantation development of in vitro fertilized zygotes developed to blastocysts in mPZM and KSOM-AA. The proportion of blastocysts developing into live pups was not different between mPZM (49%) and KSOM-AA (44%). Finally, we evaluated whether mPZM could be also used as a fertilization medium. Modified PZM containing 5.56 mM of glucose and 0.4% BSA efficiently supported IVF of mouse gametes. The percent of zygotes cleaving to 2-cell (94-98%) and blastocysts (91-93%) stage was not different from zygotes fertilized in human tubal fluid medium. We concluded that modified pig zygotic medium containing a higher potassium concentration than any other commonly used mouse media supported not only culture of mouse embryos, but also efficient IVF of mouse gametes.
authors.affiliation: School of Biosciences, University of Nottingham, Sutton Bonington Campus, Leics, UK.
authors.forename: Dasari
authors.initials: D
authors.lastname: Amarnath
authors.forename: Sayaka
authors.initials: S
authors.lastname: Wakayama
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.forename: Adel R
authors.initials: AR
authors.lastname: Moawad
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Wakayama
authors.forename: Keith H S
authors.initials: KH
authors.lastname: Campbell
mesh_terms: D000818:Animals; D001755:Blastocyst; D017077:Culture Media, Conditioned; D046149:Embryo Culture Techniques / Q000662:veterinary*; D004624:Embryo Transfer / Q000662:veterinary; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D008297:Male; D051379:Mice / Q000196:embryology*; D008815:Mice, Inbred Strains; D013552:Swine; D015053:Zygote / Q000166:cytology
chemical_list: D017077:Culture Media, Conditioned