體外胚胎自動定位與共培養微流體晶片

A Microfluidic Device for the Automatic Trapping of Embryos and the Coculture with Stromal Cells in Vitro

陳紀帆(Chi-Fan Chen)

指導教授 ： 劉承賢

國立清華大學/工學院/動力機械工程學系/碩士(2014年)

摘要

隨著工業革命、科技業興起等等人類文明的高度發展，讓現代人生活在惡劣的環境下，像是汙染、壓力、飲食等等，都是可能造成現今文明病的原因，在21世紀中，不孕的人口大大提升，生殖醫學也正致力於解決懷孕的問題，而主要治療方法多往體外受精胚胎培養發展，但提高胚胎品質的控制變因有限，且需要繁複的人工操作，因此近幾年發現微流體系統特性相當符合仿生應用，便大量發展於生殖醫學方面，而本論文研究主要為利用微機電技術與微流體理論整合自動化抓取與定位追蹤功能並應用於生醫晶片，以改良現今胚胎傳統培養方式，不但取代繁複的人工操作，更使用體外仿生環境培養，提高胚胎體外培養的品質。
重建體外仿子宮微環境，共培養胚胎與子宮內膜細胞，透過子宮內膜細胞分泌的生長因子提供養分，讓胚胎有更良好的發育環境，並藉由液壓流阻類比電路概念設計自動抓取定位結構，再以流體分析軟體ANSYS CFX最佳化流場設計，可減少人工操作對於胚胎之傷害，也便利於往後培養之觀察追蹤，培養方式則採用動態灌養培養，可排除細胞代謝之廢棄物且又可提供新鮮之培養基，為目前胚胎主要發展培養方式。
本研究論文整合自動化定位、動態灌養、共培養等多功能之體外子宮生醫晶片以提高胚胎培養品質，並於實驗中比較共培養、單養、靜態培養、動態灌養與自動定位等等不同變因之實驗，結果可發現，晶片提供良好的培養環境，有和傳統培養大致相同的囊胚比例(60% vs. 57.8%)，而生長速率方面則是較傳統方式稍快，並於實驗最後將培養之胚胎植回母體進行著床且生產出小鼠，證實本研究晶片之前瞻性與可行性。

關鍵字

共同培養 ； 體外胚胎培養 ； 生殖醫學 ； 細胞抓取

並列摘要

Infertility is a worldwide important issue. Assisted Reproductive Technology (ART) becomes one of the most important studies in the 21st Century. The reasons of infertility are most from the modern plague, such as poor nutrition, obesity, alcohol, cigarette, stress and drugs. Recently, in vitro fertilization (IVF), which holds the highest success rate, is a major treatment for infertility.
In this research, the microfluidic devices made by a simple fabrication were developed. We provided the method and operation by integrating both microfluidic techniques and coculture of stromal cells with embryos to mimic the uterus in vitro. The culturing by the dynamic system with the fresh medium also enhances the embryo development. Furthermore, the embryos were automatically and individually trapped via the flow resistance design. The design was simulated and analyzed via the commercial software, CFD and ANSYS CFX, to optimize the flow channel design as well as reduce the manual operation, which resulted in the possibility of embryos damaged. And the most important thing is that the chip is easy to track and manage individual embryos.
The integrated Labchip functions include automatic trapping, dynamic perfusion and coculture to mimic the uterus in vitro. The experimental results show that this Labchip could provide better environment compared with some different conditions such as monoculture and static culture. The embryos could be cultured in comparable blastocyst rate versus the traditional method (60% vs. 57.8%). Besides, the embryos cultured on the chip have the faster growth rate than the traditional method. All of these mean that our Labchip provides a bionic environment for embryo developing. Finally, we transfer the embryos cultured from our Labchip into the maternal uterus and born the mice. It verifies the feasibility and prospective of this Labchip.

並列關鍵字

Coculture ； In Vitro Culture ； Reproductive Technology ； Cell Trapping

參考文獻

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