New technique for mouse oocyte injection via a modified holding pipette

• Published in: Reproductive biomedicine online 2010-11, Vol.21 (5), p.663-666
• Main Authors: Lyu, Q.F, Deng, L, Xue, S.G, Cao, S.F, Liu, X.Y, Jin, W, Wu, L.Q, Kuang, Y.P
• Format: Article
• Language: English
• Subjects: Animals; Equipment Design; Fertilization in Vitro - methods; holding pipette; Humans; interspecies fertilization; intracytoplasmic sperm injection; Male; Mice; Microinjections - methods; mouse oocyte; Obstetrics and Gynecology; Oocytes; Sperm Injections, Intracytoplasmic - instrumentation; Sperm Injections, Intracytoplasmic - methods; Spermatozoa; survival rate
• ISSN: 1472-6483; 1472-6491
• DOI: 10.1016/j.rbmo.2010.07.004

Abstract To improve mouse oocyte survival from intracytoplasmic sperm injection, the sharp tip of the injection pipette has been modified to have a flat end. Here, for the same goal but for a more convenient manipulation, a sharp injection pipette was kept whereas the holding pipette was modified to have a trumpet-shaped opening, which allows deeper injection into the oocyte as it is held. Mouse oocyte injection with mouse and human spermatozoa was performed at 37°C. For the injection of mouse oocyte with mouse sperm head, a significantly higher survival rate (83%) was achieved by utilizing the modified holding pipette than the conventional one (21%; P < 0.001) and the fertilization rates were normal and comparable for both methods (82% versus 81%). A superior survival rate (82%) and acceptable normal fertilization rate (71%) were also achieved by utilizing the modified holding pipette for interspecies ICSI (injecting mouse oocyte with human spermatozoon). Taken together, by utilizing a holding pipette with a trumpet-shaped opening, acceptable rates of mouse oocyte survival and fertilization can be achieved using a sharp injection pipette under conditions usual for human oocyte injection.