Digital Microfluidic Dynamic Culture of Mammalian Embryos on an Electrowetting on Dielectric (EWOD) Chip.
Current human fertilization in vitro (IVF) bypasses the female oviduct and manually inseminates, fertilizes and cultivates embryos in a static microdrop containing appropriate chemical compounds. A microfluidic microchannel system for IVF is considered to provide an improved in-vivo-mimicking environment to enhance the development in a culture system for an embryo before implantation. We demonstrate a novel digitalized microfluidic device powered with electrowetting on a dielectric (EWOD) to culture an embryo in vitro in a single droplet in a microfluidic environment to mimic the environment in vivo for development of the embryo and to culture the embryos with good development and live births. Our results show that the dynamic culture powered with EWOD can manipulate a single droplet containing one mouse embryo and culture to the blastocyst stage. The rate of embryo cleavage to a hatching blastocyst with a dynamic culture is significantly greater than that with a traditional static culture (p<0.05). The EWOD chip enhances the culture of mouse embryos in a dynamic environment. To test the reproductive outcome of the embryos collected from an EWOD chip as a culture system, we transferred embryos to pseudo-pregnant female mice and produced live births. These results demonstrate that an EWOD-based microfluidic device is capable of culturing mammalian embryos in a microfluidic biological manner, presaging future clinical application.
Department of Obstetrics and Gynecology, Linkou Medical Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan; Department of Obstetrics and Gynecology, Chang Gung University and College of Medicine, Taoyuan, Taiwan.
Hong-Yuan
HY
Huang
Institute of Nanoengineering and Microsystem, National Tsing Hua University, Hsinchu, Taiwan.
Hsien-Hua
HH
Shen
Department of Mechanical Engineering, National Taiwan University, Taipei, Taiwan.
Chang-Hung
CH
Tien
Department of Obstetrics and Gynecology, Linkou Medical Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
Chin-Jung
CJ
Li
Department of Mechanical Engineering, National Taiwan University, Taipei, Taiwan.
Shih-Kang
SK
Fan
Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Cheng-Hsien
CH
Liu
Department of Mechanical Engineering, National Chiao Tung University, Hsinchu, Taiwan.
Wen-Syang
WS
Hsu
Institute of Nanoengineering and Microsystem, National Tsing Hua University, Hsinchu, Taiwan; Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Da-Jeng
DJ
Yao
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002470:Cell Survival; D058874:Electrowetting / Q000295:instrumentation* / Q000379:methods*; D046149:Embryo Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000502:physiology*; D005260:Female; D051379:Mice; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D011555:Pseudopregnancy