Affixing Labels on Culture Dishes Decrease the Development Rate of Mouse Embryos.

Temperature, humidity, oxygen concentration, bacteria, and airborne volatile organic compounds (VOCs) have been reported to influence the development of mammalian embryos during in vitro culturing. Identifying and managing culture embryos, we are able to use a pen or labels for writing patient's information on the plastic ware. There has been concern that the VOCs originated from the ink of felt-tip pens, which are too small to be removed with the HEPA filter, might cause the decrease of the developmental rate of culture embryos. On the other hand, verification report is less about the labels. This study was conducted to investigate effects of affixing labels on culture dishes on development of the embryos in the mouse. Prospective experimental Animal study. We performed ovarian hyperstimulation on 8-week-old female BDF1 mice by administering eCG 7.5 IU or hCG 7.5 IU. After rearing these females in the same cage with males of the BDF1, fertilized eggs were collected from oviducts of the plug-positive females. Plastic dishes (Falcon) with a 35-mm aperture were prepared with five 20-μL drops of M16, covered with mineral oil (Vitrolife), placed in a 60-mm dish (Falcon), and then cultured at rest in an incubator (HC-3100 ASTEC). Five to ten embryos were cultured per drop. Labels using acrylic adhesive were affixed to the inside of the lid of the 60-mm dish, and culturing was performed at 37°C, 5% CO2, 5% O2, 90% N2, and saturated humidity. The number of affixed labels totaled 0 (control group), 1 (group A), 7 (group B), or 13 (group C), and development of the cultured mouse embryos from the 1-cell stage was investigated for 96 hours. Statistically analysis between experimental groups were determined by one-way ANOVA followed by Tukey test for multiple comparisons. Following the approval by the Animal Care and Use Committee of Azabu University, we used 263 mouse embryos in this study, which was performed at the Azabu University in the duration from September 2015 to December 2015. The cleavage rate of the control group and groups A, B, and C were 97.5%, 97.1%, 12.0%, and 9.5%, respectively, with significant (P<0.001) decreases in groups B and C compared with the control; the developmental rate to the blastocyst stage were 88.6%, 0%, 0%, and 0%. As increasing the number of label, the incidences of cleavage of the cultured embryos were reduced, and there was no embryo developed to the blastocyst stage in all of the label-affixed groups. Affixing labels using acrylic adhesive on culture dishes have detrimental effects on the development of mouse embryos at 1-cell stage. It is concluded that careful consideration should be given to the adhesive's components when using label stickers for culture dish management.