Huang Hong-Yuan, Lai Yun-Li, Yao Da-Jeng
Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, 5, Fu-Hsing Street, Kwei-Shan, Tao-Yuan 333, Taiwan.
Department of Obstetrics and Gynecology, Chang Gung University and College of Medicine, 259, Wen-Hua 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan.
Micromachines (Basel). 2018 Mar 20;9(3):135. doi: 10.3390/mi9030135.
The aim of this work was to create a microfluidic platform that uses in vitro fertilization (IVF) and avoids unnecessary damage to oocytes due to the dielectrophoretic force manipulation of the sperms and oocytes that occurs in a traditional IVF operation. The device from this research can serve also to decrease medium volumes, as well as the cost of cell culture under evaporation, and to prevent unnecessary risk in intracytoplasmic sperm injection (ICSI). To decrease the impact and destruction of the oocyte and the sperm, we adopted a positive dielectrophoretic force to manipulate both the sperms and the oocyte. The mouse oocytes were trapped with a positive dielectrophoretic (p-DEP) force by using Indium Tin Oxide (ITO)-glass electrodes; the ITO-glass electrode chip was fabricated by wet etching the ITO-glass. The polydimethylsiloxane (PDMS) flow-focusing microfluidic device was used to generate microdroplets of micrometer size to contain the zygotes. The volume of the microdroplets was controlled by adjusting the flow rates of both inlets for oil and the DEP buffer. As a result, the rate of fertilization was increased by about 5% beyond that of the DEP treatment in traditional IVF, and more than 20% developed to the blastocyst stage with a low sperm-oocyte ratio.
这项工作的目的是创建一个微流控平台,该平台采用体外受精(IVF)技术,避免传统IVF操作中因精子和卵母细胞的介电泳力操纵而对卵母细胞造成不必要的损伤。本研究中的装置还可用于减少培养基体积,以及蒸发条件下细胞培养的成本,并防止胞浆内单精子注射(ICSI)中的不必要风险。为了减少对卵母细胞和精子的影响及破坏,我们采用正介电泳力来操纵精子和卵母细胞。通过使用氧化铟锡(ITO)玻璃电极,利用正介电泳(p-DEP)力捕获小鼠卵母细胞;ITO玻璃电极芯片通过对ITO玻璃进行湿法蚀刻制成。聚二甲基硅氧烷(PDMS)流动聚焦微流控装置用于生成微米级的微滴以容纳受精卵。通过调节油和DEP缓冲液两个入口的流速来控制微滴的体积。结果,受精率比传统IVF中的DEP处理提高了约5%,并且在低精子-卵母细胞比例下超过20%发育到囊胚阶段。