Evaluation of Effect of Hydrodynamic Shear Stress on Cultured Plant Cells

水动力剪切应力对培养植物细胞影响的评价

• 批准号: 07555255
• 负责人: FURUSAKI Shintaro
• 金额: $ 3.46万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07555255/
• 关键词: Plant Cell Culture; Hydrodynamic Forces; Shear Stress; Stirred Tank; Viability; Aggregate Size; TTC; Power Input; シアストス; 死滅速度

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.

本研究的目的是探讨利用有限而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.植物生活力测定：在细胞受到剪切应力后，用染料试验和TTC试验检测样品的活力。结果强烈建议TTC测定植物活力应仅用于定性试验2.层流剪切应力对植物细胞存活力的影响：该装置被制造成在植物细胞上经受不同水平的层流剪切应力。在间歇培养中，对数生长期和稳定生长期对剪切应力的抵抗能力最强，接种时间对剪切应力的抵抗能力最强。3.湍流水动力对植物细胞的影响：1）在层流应力装置中，不同搅拌器的临界输入功率与临界剪切应力之间存在着一定的相关性。结果表明，尽管植物细胞和无机模型系统的成熟度不同，但它们的数据有很多共同之处。因此，无机系统能够用于研究以确定搅拌容器中的流体动力。2)在胁迫细胞的胞质溶胶中发现溶酶体的关键酶（酸性磷酸酶），浓度略高于对照组。3)不是在整个生物反应器中耗散的功率，而是直接在搅拌器周围耗散的体积与细胞的耗氧速率具有良好的相关性。4)粉红色念珠菌细胞需要水动力来生产阿马里辛。5)尽管平均功率输入相同且局部耗散功率输入较低，但与Rushton涡轮机相比，锚式搅拌器产生的聚集体较小。