仿肝小葉與肝腺泡之微流體細胞培養晶片

Microfluidic cell-culturing Devices Mimicking Liver Lobule and Acinus

石明正(Ming-Cheng Shih)

指導教授 ： 劉承賢

國立清華大學/工學院/奈米工程與微系統研究所/博士(2013年)

https://doi.org/10.6843/NTHU.2013.00417

摘要

本論文的研究目的是希望設計生物反應器能維持rat primary liver cell CYP450活性，希望在反應器中藥物測試呈現有藥劑的濃度的相關影響並與動物體內的呈現一致的結果。
本論文的研究分成兩個部分，第一個部分為組合式的MTR(Micro Tissue Reactor)，主要以模擬體內liver loube內特殊的流場結構，提供體外liver cell所需的microenviroment，達到能維持rat primary liver cell CYP450活性，並在反應器中藥物測試呈現有藥劑的濃度的相關影響並與動物體內的呈現一致的趨勢。
第二部分的acinus chip的設計是為了彌補MTR功能的不足，希望能在cell seeding後馬上開始perfusion，不用因為等待細胞貼附的時間，導致CYP450活性快速下降，並且提供cell membrane directly contact的狀態使得能夠進行cell-cell interaction，同時能動態調整或觀測acinus-like gradient的變化，調控shear stress和mass-transfer達到平衡，使體外細胞培養的microenviroment更趨近於體內，維持體外liver細胞解毒功能時間更長，以便於用於藥物毒性篩選。為了方便操作多個acinus chip 進行生物實驗，因此設計了 96 well compatible format，搭配PC通訊控制晶片內medium流動的速度，整合成一套可即時觀測的細胞培養系統。

關鍵字

微流體 ； 肝細胞 ； 肝小葉 ； 肝腺泡 ； 細胞培養

並列摘要

The objective of this research was to design a micro-bioreactor to sustain the CYP450 activity in the primary rat liver cells and to understand the related impact by the concentration of the applied medications. The study was divided into two parts. The first part involved a micro-tissue reactor (MTR) which primarily simulated the specific flow field structure of the liver lobule to provide a similar microenvironment to the liver cells outside the body. The purpose was to sustain and observe the effect of drug concentrations on the CYP450 activity in the primary rat liver cells under an in vitro environment that is comparable to the true physiological conditions.
In the second part, the design of the acinus chip was to compensate for the inadequacies of the MTR such as cell adherence. The process of cell adherence usually ends with rapid degradation of the CYP450 activity. This new design would give the researchers the option of immediate perfusion after cell seeding without waiting for cell adherence. Also, the acinus chip could provide direct contact with the cell membrane which allows researchers to observe cell-to-cell interactions. In addition to providing the feature of dynamic control for observing the acinus-like gradient change, the acinus chip could also control and balance the shear stress and mass-transfer of the microenvironment. The end result is an in vitro microenvironment that closely resembled the true physiological state. The design allowed liver cells to maintain cytotoxic detoxification function even outside the body, and thus, made drug toxicity studies more convenient.

並列關鍵字

microfluidic ； liver cell ； liver lobule ； liver acinus ； cell culture

參考文獻

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