Cell Damage due to Osmotic Stress and Microscale Mass Transfer relative to Cryopreservation

渗透应力和与冷冻保存相关的微量传质引起的细胞损伤

• 批准号: 15360116
• 负责人: ISHIGURO Hiroshi
• 金额: $ 8.64万
• 依托单位: Kyusyu Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15360116/
• 关键词: Cryopreservation; Cell damage; Electrolyte solution; Solution effect; Concentration; Exposure time; Temperature; Mathematical model; 細胞; 生存率

In this study, cell damage and death due to the hypertonic electrolyte solution under the unfrozen condition were investigated experimentally in relation to the potential causes of freezing damage of cells. Next, a mathematical model with a form of reaction kinetics was proposed and developed for the cell damage and death.1) The cells from a human prostatic adenocarcinoma cell line (PC-3) were exposed from an isotonic solution (physiological saline) to hypertonic electrolyte solution (NaCl concentration C_m and exposure time τ) back to the isotonic solution under a constant temperature, T℃. Then, the cell membrane integrity was assessed in physiological saline at 25℃ to measure the cell viability. T was set at three values between 4 and 25℃, C_m at four values between 1.8 and 4.0M, and τ at seventeen values between 2 and 170min.2) From the experimental results, survival curve of cell as a function of τ, probability distribution and probability density function of cell death, characteri … More stic exposure times of cell death, etc. were obtained to clarify the dependency of them on τ, C_m, and T.3) The cell viability decreased with increasing exposure time. The decrease in viability was steeper at a higher concentration and higher temperature. When a characteristic exposure time for the decrease in cell viability was defined as exposure time τ_<0.5> at 0.5 in cell viability, the viability versus nondimensional exposure time τ/τ_<0.5> had a similar distribution independently of concentration and temperature. This means that the cell damage and death proceed kinetically and similarly, suggesting that the mechanism may be also similar.4) The cell damage and death process was formulated with a kinetic model of two-step damage on the basis of experimental results of the cell viability. Here, three states of cells : vigorous, damaged but living (medium state), and dead were considered. Four kinds of model were proposed and investigated. A reaction rate constant in the model was determined by the inverse problem analysis and the best model of four was determined. The cell viability was simulated successfully by the analytical solution from the present model. Also, the probability distribution and probability density function of cell death, and reaction kinetic properties were obtained. The reaction kinetic constant increased with an increase in C_m and T. Less

在这项研究中，通过实验研究了非冷冻条件下高渗电解质溶液造成的细胞损伤和死亡，并与细胞冷冻损伤的潜在原因相关。接下来，提出并开发了一种反应动力学形式的数学模型。 1)将人前列腺腺癌细胞系(PC-3)的细胞从等渗溶液(生理盐水)暴露于高渗电解质溶液(NaCl浓度C_m和暴露时间) τ）回到恒温T℃下的等渗溶液，然后在25℃的生理盐水中评估细胞膜的完整性，以测量细胞活力，T设定在4至25℃之间的三个值。 1.8到4.0M之间有四个值，2到170min之间有十七个值。2）从实验结果来看，细胞的生存曲线作为τ的函数，细胞的概率分布和概率密度函数获得了细胞死亡的死亡、特征暴露时间等，以阐明它们对 τ、C_m 和 T 的依赖性。3) 细胞活力随着暴露时间的增加而下降，活力的下降在较高的情况下更加陡峭。当细胞活力下降的特征暴露时间定义为细胞活力为 0.5 时的暴露时间 τ_<0.5> 时，活力与无量纲暴露时间 τ/τ_<0.5> 具有相似的分布，独立于细胞活力。这说明细胞损伤和死亡在动力学上是相似的，提示其机制可能也是相似的。4)在实验的基础上，建立了细胞损伤和死亡过程的两步损伤动力学模型。这里，考虑了细胞的三种状态：活跃的、受损但存活的（中等状态）和死亡的模型，并通过反演问题确定了模型中的反应速率常数。分析并确定了四个最佳模型的细胞活力。通过该模型的解析解成功地模拟了细胞死亡的概率分布和概率密度函数，以及反应动力学常数随着C_m和T的增加而增加。

项目成果

Cell Viability and Kinetic Model for Prediction of Viability under Exposure to Hypertonic NaCl Solution at Near-zero Temperatures (2nd)

用于预测在接近零温度下暴露于高渗 NaCl 溶液时的活力的细胞活力和动力学模型（第二）

• 发表时间: 2006
• 作者: Hiroshi ISHIGURO; et al.
• 通讯作者: et al.

Cell Viability and Kinetic Model for Prediction of Viability under Exposure to Hypertonic NaCl Solution at Near-zero Temperatures (2nd)

用于预测在接近零温度下暴露于高渗 NaCl 溶液时的活力的细胞活力和动力学模型（第二）

• 发表时间: 2006
• 作者: Hiroshi ISHIGURO; et al.
• 通讯作者: et al.

Reaction Kinetic Modeling of Cell Damage and Death due to Hypertonic Electrolyte Solution

高渗电解质溶液引起的细胞损伤和死亡的反应动力学模型

• 发表时间: 2005
• 作者: Hiroshi ISHIGURO; et al.
• 通讯作者: et al.

高張電解質水溶液による細胞損傷・死滅の相似的特性(非凍結低温条件)

高渗电解质溶液（非冰冻低温条件）引起细胞损伤和死亡的相似特征

• 发表时间: 2005
• 作者: 石黒 博; 他1名
• 通讯作者: 他1名

高張電解質水溶液による細胞損傷・死滅の反応速度論的モデルの展開

高渗电解质溶液引起的细胞损伤和死亡的动力学模型的建立

• 发表时间: 2005
• 作者: 石黒 博; 他1名
• 通讯作者: 他1名