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当悬浮液进行离心时影响细胞产量和存活率的因素。在准均匀离心场中离心加速度、离心时间和悬浮液柱长度的影响。

Factors affecting yield and survival of cells when suspensions are subjected to centrifugation. Influence of centrifugal acceleration, time of centrifugation, and length of the suspension column in quasi-homogeneous centrifugal fields.

作者信息

Katkov I I, Mazur P

机构信息

Semen and Embryo Preservation Laboratory, Universal Preservation Technologies, Inc., San Diego 92121, USA.

出版信息

Cell Biochem Biophys. 1999;31(3):231-45. doi: 10.1007/BF02738241.

Abstract

The goals of the centrifugation of cell suspensions are to obtain the maximum yield of cells with minimum adverse effects of centrifugation. In the case of mechanically sensitive cells such as mouse sperm, the two goals are somewhat contradictory in that g-forces sufficient to achieve high yields are damaging, and g-forces that yield high viability produce low yields. This paper mathematically analyzes the factors contributing to each goal. The total yield of pelleted cells is determined by the sedimentation rate governed by Stokes' Law, and depends on the relative centrifugal force, centrifugation time, size and shape of the cells, density of the cells and medium, viscosity of the medium, and the length of the column of suspension. Because in the situation analyzed the column is short relative to the rotor radius, the analysis considers the centrifugal field to be quasi-homogeneous. The assumption is that cells are not damaged during sedimentation, but that they become injured at an exponential rate once they are pelleted, a rate that will depend on the specific cell type. The behavior is modeled by the solution of coupled differential equations. The predictions of the analysis are in good agreement with experimental data on the centrifugation of mouse sperm.

摘要

细胞悬液离心的目标是在使离心的不利影响最小化的情况下获得最大产量的细胞。对于像小鼠精子这样对机械敏感的细胞而言,这两个目标在某种程度上是相互矛盾的,因为足以实现高产率的重力会造成损害,而能产生高活力的重力则会导致低产率。本文对影响每个目标的因素进行了数学分析。沉淀细胞的总产量由斯托克斯定律所支配的沉降速率决定,并取决于相对离心力、离心时间、细胞的大小和形状、细胞与培养基的密度、培养基的粘度以及悬浮液柱的长度。由于在所分析的情况下,柱相对于转子半径较短,所以该分析将离心场视为准均匀的。假设是细胞在沉降过程中不会受损,但一旦沉淀下来就会以指数速率受到损伤,该速率将取决于特定的细胞类型。这种行为通过耦合微分方程的解来建模。分析的预测结果与小鼠精子离心的实验数据高度吻合。

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