Evaluation of Effect of Hydrodynamic Shear Stress on Cultured Plant Cells

本研究旨在探讨利用有限和简单的实验结果来预测细胞在强水动力胁迫下的生长和生产行为的可行性。1.植物活力测定：细胞受到剪切胁迫后，用染料试验和TTC法检测细胞的活力。结果表明，TTC法测定植物活力只应作为定性测试。2.层流剪应力对植物细胞活力的影响：该装置是为了对植物细胞施加不同程度的层流剪应力而制作的。在间歇培养中，指数生长和平稳生长之间的阶段对剪应力的抵抗能力最强，接种时间最敏感。3.湍流水动力对植物细胞的影响：1)具有不同类型搅拌器的搅拌槽中的临界功率输入值与层流应力装置中的临界应力值相关联。结果表明，植物细胞和无机模型系统的数据显示出许多相似之处，尽管它们的成熟度截然不同。因此，无机体系能够进行研究，以确定搅拌容器中的水动力。2)与对照相比，胁迫细胞胞浆中的溶酶关键酶(酸性磷酸酶)的浓度略高。3)不是在整个生物反应器内消耗的能量，而是直接在搅拌器周围耗散的体积与细胞的耗氧率有很好的相关性。4)玫瑰花细胞需要水动力才能产生阿马利星。5)尽管锚定搅拌器的平均功率相同且局部耗散功率较低，但与Rushton水轮机相比，锚定搅拌器产生的聚集体较小。

The purose of this study is to investigate the feasibility of using results from limited and easy experiments for predicting for predicting the cell's growth and production behavior under intense hydrodynamic stress.1.Plant viability assay : After cells were stressed with shear, samples were examined for viability with the dye test as well as TTC.The results strongly suggests TTC assay for plant viability determination should be used only as qualitative test.2.Influence of laminar shear stress upon plant cells' viability : The device was produced to subject different levels of laminar shear stress on plant cells. The phase between the exponential and stationary growth revealed to be the most resistant to shear stress in batch culture and the time of inoculation the most sensitive.3.Influence of turbulent hydrodynamic forces upon plant cells : 1) The critical power input values in a stirred tank with different types of stirrer were correlated with the critical sheat stress values in the laminar stress device. The results show that the data found for plant cells and for the inorganic model system showed much in common despite their distinct different mature. Therefore the inorganic system is capable for investigations to determine hydrodynamic forces in agitated vessels. 2) Key enzymes of lysosmes (acid phosphatase) were found in the cytosol of stressed cells in slightly higher concentration compared to the control. 3) Not the power dissipated in the whole bioreactor but dissipated the volume directly around the stirrers gives good correlation with oxygen consumption rate of cells. 4) C.roseus cells required hydrodynamic forces for ajmalicine production. 5) In spite of the same mean and lower locally dissipated power input, the anchor stirrers generated smaller aggregates compared to the Rushton Turbines.