A Microfluidic Device for Automatic Trapping and Co-culture of Mice Embryos with Human Endometrial Stromal Cell
| Autor: | Sung, Yi-jung, 宋宜蓉 |
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| Rok vydání: | 2016 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 104 Because of the high education, rapid urbanization, enhancement of feminism, financial independence of women and fluctuating economy in Taiwan, most of people marry late. Hence, population of fertility in advanced age grows obviously. The fertility rate would decrease as marriage age. Except for age factors, DNA damage, genetic factor, abnormality of reproductive system and hormone imbalance also result in infertility. The mainstream therapy is in-vitro fertilization (IVF), namely test-tube baby. This method requires complicated operating sequences and well-trained technicians. The quality of IVF is also limited. According to statistical data in 2011, The average fertility rate of IVF is about 27.4% in Taiwan. How to enhance the embryo quality is a crucial target for IVF studies in the future. The microfluidics could reach lots of bionic applications. Therefore, recent research tends to develop microfluidics for various biomedical applications. In this study, we developed microfluidic Labchip system combining automatic trapping and dynamic culture system to improve traditional IVF method for reproductive biology application. In addition to replacing complicated artificial operation, a bionic culture environment was developed in this research to enhance embryo quality. In this research, we co-cultured embryos with endometrial stromal cells and used dynamic perfusion system with porous membrane to build in-vitro uterus environment by the means of growth factors which stromal cells secrets to provide a uterus-mimicking growth environment for embryonic development. Dynamic culture can not only carry metabolic waste off but also supply fresh medium to maintain stable osmotic pressure and pH value. We also implemented to the automatic trapping system via fluidic resistance design and manipulation to reduce embryo damage resulting from delicate artificial operation. According to the experimental results, we successfully captured embryos by adjusting the height of channel to 140μm. The stromal cells grew faster in porous membrane and reached to 70% after 2.5 days culturing. We compared monoculture and coculture in petri dish after culturing 2.5 days. The blastocyst developing rate (BDR) of monoculture is 15.79%, and the BDR of cocultrue is 57.89%. It means coculure with stromal cells grew much better and faster. In comparison with monoculture in chip and petri dish, BDR of the former is 57.10% and BDR the latter is 57.58%. Therefore, there is no significant difference between culturing in chip or in petri dish. We compared the coculture in chip and in petri dish. The BDR of coculture in chip is 58.33%, and the BDR of cocultrue in petri dish is 26.67%. Coculture in petri dish is not able to work. On the other hand, coculture in chip via the microfluidic system did successfully work. The embryo transfer rate is 66.67%. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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