Culture of mouse preimplantation embryos as a quality control assay for human in vitro fertilization

Abstract

Techniques for culturing preimplantation mouse embryos from the two-cell stage to blastocyst are described, and the importance of this system to quality control assay the media and supplements used in human in vitro fertilization (IVF) procedures is discussed. Embryos from B6CBAF, mice were cultured in a commonly used mouse culture medium, modified Krebs-Ringer-bicarbonate (Krebs'), or in a commonly used human culture medium, Ham's FIO nutrient mixture supplemented with human fetal cord serum (FCS), and results were not significantly different. Using the mouse embryo culture system, tests on 174 preparations of FCS resulted in 24.1 % producing less than 75% morula or blastocyst stages after 72 h in culture, compared to 9.5% of the Krebs' control cultures. Rcsults of the mouse embryo culture system using 98 FCS subsequently used in human IVF were compared to results from the VIP Human In Vitro Program of Eastern Virginia Medical School of Norfolk, Virginia, from June 1982 through June 1983. The data suggest that prescreening of media and supplements using this mouse embryo culture system may indicate sources of factors potentially detrimental to the success of human IVF procedures.

Double-quality control reveals high-level toxicity in gloves used for operator protection in assisted reproductive technology

“…Several studies (1,2,(4)(5)(6)(7)(8)(9)(10)(11)(16)(17)(18) have shown that successful culture of human embryos in an IVF program requires good laboratory procedures and selection of nonembryo-toxic equipment and culture products. Different embryonic stages show different sensitivities to toxic effects, and this is why we have used both (ZPþ) and (ZP-) zygotes and not only fresh but also frozen/thawed embryos (1,4,10).…”

Characterization of secreted proteins of 2-cell mouse embryos cultured in vitro to the blastocyst stage with and without protein supplementation

“…Here we cultured 2-cell embryos to the blastocyst stage; the observed development and proportions of blastocysts reaching expanded and hatching stages were similar under culture conditions with and without protein supplementation. We have used this system for embryo toxicity and quality control in our IVF program for several years [19]. We pooled samples and used concentration methods in order to increase protein abundances, and embryos were cultured without change in media for the 3-day period in an effort to accumulate secretory products.…”

Quality control measures in an embryo research program

“…Therefore, an alternative model like the rabbit (Kuehl et al 1986) may be useful Details for establishing mouse embryo cultures for quality control purposes appear elsewhere (Hogan et al, 1986;Monk, 1987;Gerrity, 1988). For human in vitro fertilization programs, 1-to 4-cell mouse embryos are used, and development is monitored to the hatched b astocyst stage over a 3-to 5-day period (Ackerman et al, 1984;Trouson andConto 1984-Heming et al, 1987). Cryopreserved 2-cell mouse embryos can be purchased from commercial sources for quahty control assessments (Leibo, 1990).…”