Dept. of Chemical and Biomedical Engineering, Cleveland State University, Cleveland, OH 44115, USA.
Biotechnol Prog. 2010 Sep-Oct;26(5):1361-6. doi: 10.1002/btpr.432.
Accurate cell settling velocity determination is critical for perfusion culture using a gravity settler for cell retention. We have developed a simple apparatus (a "settling column") for measuring settling velocity and have validated the procedure with 15-μm polystyrene particles with known physical properties. The measured settling velocity of the polystyrene particles is within 4% of the value obtained using the traditional Stokes' law approach. The settling velocities of three hybridoma cell lines were measured, resulting in up to twofold variation among cell lines, and the values decreased as the cell culture aged. The settling velocities of the nonviable cells were 33-50% less than the corresponding viable cells. The significant variation of settling velocities among cell populations and growth phases confirms the necessity of routine measurement of this property during long-term perfusion culture.
准确测定细胞沉降速度对于使用重力沉降器进行细胞保留的灌注培养至关重要。我们开发了一种简单的装置(“沉降柱”)来测量沉降速度,并使用具有已知物理特性的 15μm 聚苯乙烯颗粒对该程序进行了验证。聚苯乙烯颗粒的测量沉降速度与使用传统斯托克斯定律方法获得的值相差在 4%以内。测量了三种杂交瘤细胞系的沉降速度,结果表明细胞系之间存在两倍的变化,并且随着细胞培养老化,沉降速度降低。死亡细胞的沉降速度比相应的活细胞低 33-50%。细胞群体和生长阶段的沉降速度的显著变化证实了在长期灌注培养过程中常规测量该特性的必要性。