Bierau H, Perani A, al-Rubeai M, Emery A N
Centre for Bioprocess Engineering, School of Chemical Engineering, University of Birmingham, Edgbaston, UK.
J Biotechnol. 1998 Jul 16;62(3):195-207. doi: 10.1016/s0168-1656(98)00064-9.
It is demonstrated, using two different perfusion reaction systems, that hybridoma modified by inhibiting their apoptotic response can give improved process performance in terms of cell number and viability in intensive cell culture. Two cell perfusion systems, one using a spin filter and the other an ultrasonic filter, are compared using two cell lines. One cell line is transfected with the bcl-2 gene (TB/C3 bcl-2) which encodes the 'anti-apoptotic' human bcl-2 protein and the other cell line (TB/C3 pEF) with a negative transfection vector. Both reactor systems give similar retention performance for both cell lines. Bcl-2 transfected cells reach higher cell densities than the control cell line, and the percentage of apoptotic cells is clearly lower than with pEF cells. The maximum cell numbers of the bcl-2 cell line are 1.21 x 10(7) ml-1 in the ultrasonic filter culture and 1.58 x 10(7) ml-1 in the spin filter culture, respectively. Using the pEF cell line the maximum cell number reaches 6.0 x 10(6) ml-1 with ultrasonic retention and 5.9 x 10(6) ml-1 in the spin filter. The use of ultrasound in this cell retention system has no apparent influence on cell growth, productivity or viability. Selective retention of viable cells is detectable but the effect of removing non-viable cells is negligible.
使用两种不同的灌注反应系统证明，通过抑制其凋亡反应进行修饰的杂交瘤在密集细胞培养中的细胞数量和活力方面可以提高工艺性能。使用两种细胞系比较了两种细胞灌注系统，一种使用旋转过滤器，另一种使用超声过滤器。一种细胞系用编码“抗凋亡”人bcl-2蛋白的bcl-2基因（TB/C3 bcl-2）转染，另一种细胞系（TB/C3 pEF）用阴性转染载体转染。两种反应器系统对两种细胞系都具有相似的保留性能。bcl-2转染细胞的细胞密度高于对照细胞系，并且凋亡细胞的百分比明显低于pEF细胞。bcl-2细胞系在超声过滤器培养中的最大细胞数分别为1.21×10⁷ ml⁻¹，在旋转过滤器培养中为1.58×10⁷ ml⁻¹。使用pEF细胞系，超声保留时的最大细胞数达到6.0×10⁶ ml⁻¹，在旋转过滤器中为5.9×10⁶ ml⁻¹。在这种细胞保留系统中使用超声对细胞生长、生产力或活力没有明显影响。可检测到活细胞的选择性保留，但去除非活细胞的效果可忽略不计。
