New technique for mouse oocyte injection via a modified holding pipette.
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.
Lyu
Q F
QF
The Department of Assisted Reproductive Technology, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China.
Deng
L
L
Xue
S G
SG
Cao
S F
SF
Liu
X Y
XY
Jin
W
W
Wu
L Q
LQ
Kuang
Y P
YP
D000818:Animals; D004867:Equipment Design; D005307:Fertilization in Vitro / Q000379:methods*; D006801:Humans; D008297:Male; D051379:Mice; D008845:Microinjections / Q000379:methods; D009865:Oocytes; D020554:Sperm Injections, Intracytoplasmic / Q000295:instrumentation* / Q000379:methods; D013094:Spermatozoa