Hypoxanthine causes a 2-cell block in random-bred mouse embryos.
Ham's F-10, a chemically defined, complex culture medium, commonly used for in vitro fertilization of human as well as animal oocytes, blocked development at the 2-cell stage of greater than 92% of embryos from random-bred Swiss mice (CD-1), but did not block development of embryos from hybrid-inbred mice (BDF1). In contrast, BWW, a simple, modified Kreb's-Ringer bicarbonate medium, supported development to blastocysts of 85% and 100% of 2-cell embryos from CD1 and BDF1 females, respectively. As little as 15% (v/v) Ham's F-10 added to the BWW blocked the development of the random-bred embryos. Supplementing the BWW with Ham's F-10 components revealed that hypoxanthine (6-30 microM) was responsible for the developmental block to the random-bred embryos. The hypoxanthine block was partially (40%) reversed by adding the chelating agent, ethylenediaminetetraacetic acid. Breeding experiments showed that the hypoxanthine sensitivity of embryos from CD-1 mothers was not affected by the paternal genome.
Loutradis
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Department of Obstetrics, Harvard Medical School, Boston, Massachusetts 02115.
John
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Kiessling
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D000818:Animals; D003470:Culture Media; D004622:Embryo, Mammalian / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology*; D051379:Mice; D008815:Mice, Inbred Strains; D009924:Organ Culture Techniques
D003470:Culture Media; D007042:Hypoxanthines; D019271:Hypoxanthine