PC12細胞株不僅具有類似神經細胞的型態，而且兼具細胞內氧含量下降之反應特性，因此廣泛地應用在細胞外的缺氧性中風機制研究。本研究的目的主要是發展一套適合PC12細胞觀察應用之即時培養條件監控測系統，和探討在沒有染色的情況下找出有用的參數以分辦細胞的死亡型態；從實驗結果中顯示，本研究所建構的培養系統，在溫度的控制方面可達到37.1 ± 0.5℃，平均相對溼度方面可達到95%，氧氣濃度(5%)的監測可控制在4.88 ± 0.3%，及二氧化碳濃度(5%)的測試值為5.72 ± 0.5%。至於細胞影像分析方面，本研究實現與應用特徵分析參數如核質比、面積、熵值和平均灰階值等參數去量化細胞膜和細胞核；根據單一細胞的分析結果顯示，細胞的凋亡和計畫性死亡型態於實驗開始後13至19個小時期間，可以由一正規化後的域值來區分，分別為膜的熵值(0.29)，膜的平均灰階值(0.26)，核的面積(0.16)，細胞核質比(0.14)。以多細胞的實驗結果顯示，低缺氧(1%)環境對於細胞的傷害大於中缺氧(5%)環境，無萄葡糖培養基和中缺氧的情形比有萄葡糖培養基和中缺氧的情形傷害來得大。

The PC12 cell line has been widely used in stroke related in vitro study, due to its features of neuronal-like morphology and cell-like phenomenon under O2 deprived condition. The purposes of this study are to develop a real-time culturing conditions monitoring system for PC12 application, and to investigate the useful parameters for characterizing the types of cell death with no immunoassay labeling. From the experimental results, it is shown that the temperature, the mean relative humidity, the O2 concentration (5%), and the CO2 concentration (5%) for the proposed culturing system are controlled to be 37.1 ± 0.5℃, 95%, 4.88 ± 0.3%, and 5.72 ± 0.5%, respectively. For the cell image analysis, the feature parameters of the N/C ratio, area, and entropy are realized and applied to quantify the membrane and nucleus of PC12. From the experimental results of single cell continuously monitoring, the apoptosis or necrosis of a cell is differentiated using the thresholds of parameters of the entropy (0.29) and mean gray value (0.26) of membrane, area of nucleus (0.16), and N/C ratio (0.14) after being cultured for 13 to 19 hours. From the experimental results of multiple cells observations, it is shown that the OGD (5% O2) and lower hypoxic (1% O2) conditions would damage the cells more than that of mild hypoxic (5% O2) conditions.

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