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哺乳动物细胞培养悬浮液的流变性:杂交瘤和 HeLa 细胞系。

Rheological properties of mammalian cell culture suspensions: Hybridoma and HeLa cell lines.

机构信息

Department of Chemical Engineering, University of California at Davis, California 95616, USA.

出版信息

Biotechnol Bioeng. 1993 Mar 25;41(7):745-54. doi: 10.1002/bit.260410709.

Abstract

Data on viscous (eta') and elastic (eta'') components of the complex viscosity versus oscillatory angular frequency (0.01 to 4.0 rad/s) with increasing strains were obtained for hybridoma cell (62'D3) and HeLa cell (S3) suspensions in PBS at 0.9 (mL/mL) cell volume fraction using a Weissenberg rheogoniometer equipped with two parallel plate geometry at ambient temperature. Both cell suspensions exhibited shear thinning behavior. From the measured viscoelastic properties, the yield stress was calculated. Hybridoma cell suspension (15 microm as the mean diameter of cells) showed the yield stress at 550 dyne/cm(2) that was 1.8 times higher than the value of HeLa cell suspension (22 microm mean diameter) as measured at the oscillatory angular frequency, 4.0 rad/s. The apparent viscosities of HeLa cell suspension at four concentrations and varying steady shear rate were also determined using the Brookfield rotational viscometer. The yield stress to steady shear test was about 130 dyne/cm(2) for HeLa cell suspension at 0.9 (mL/mL) cell volume fraction. The apparent viscosity was in the range about 1 approximately 1000 Poise depending on the cell concentration and shear rate applied. A modified semiempirical Mooney equation, eta = eta(0) exp[K gamma(.)(-beta)phi(c)(1 - K'' sigmaphi(c) /D)] was derived based on the cell concentration, the cell morphology, and the steady shear rate. The beta, shear rate index, was estimated as 0.159 in the range of shear rate, 0.16 to 22.1 s(-1), for the cell volume fractions from 0.6 to 0.9 (mL/mL). In this study, the methods of determining the shear sensitivity and the viscous and the elastic components of mammalian cell suspensions are described under the steady shear field.

摘要

在环境温度下,使用配备有平行板几何形状的 Weissenberg 流变仪,在 PBS 中以 0.9(mL/mL)细胞体积分数获得杂交瘤细胞(62'D3)和 HeLa 细胞(S3)悬浮液的复数粘度的粘性(eta')和弹性(eta'')分量与振荡角频率(0.01 至 4.0 rad/s)的关系数据。在振荡角频率为 4.0 rad/s 时,两种细胞悬浮液均表现出剪切稀化行为。从测量的粘弹性特性中计算出屈服应力。杂交瘤细胞悬浮液(细胞平均直径为 15 微米)的屈服应力为 550 dyne/cm(2),比 HeLa 细胞悬浮液(细胞平均直径为 22 微米)的屈服应力高 1.8 倍。使用 Brookfield 旋转粘度计还确定了四种浓度和变化的稳态剪切速率下的 HeLa 细胞悬浮液的表观粘度。在 0.9(mL/mL)细胞体积分数下,HeLa 细胞悬浮液的屈服应力到稳态剪切测试约为 130 dyne/cm(2)。表观粘度取决于细胞浓度和施加的剪切速率,范围约为 1 至 1000 泊。基于细胞浓度、细胞形态和稳态剪切速率,推导出了一个改进的半经验 Mooney 方程,eta = eta(0) exp[K gamma(.)(-beta)phi(c)(1 - K'' sigmaphi(c) /D)]。在剪切速率为 0.16 至 22.1 s(-1)、细胞体积分数为 0.6 至 0.9(mL/mL)的范围内,估计剪切率指数 beta 为 0.159。在本研究中,描述了在稳态剪切场下确定哺乳动物细胞悬浮液剪切敏感性以及粘性和弹性分量的方法。

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