Mollet Mike, Godoy-Silva Ruben, Berdugo Claudia, Chalmers Jeffrey J
Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W 19th Ave., Columbus, Ohio 43210, USA.
Biotechnol Bioeng. 2007 Nov 1;98(4):772-88. doi: 10.1002/bit.21476.
A second generation flow contraction device was developed and modeled which allows cells to be subjected to well-defined hydrodynamic forces. Studies were conducted with this system on wild-type Chinese Hamster Ovary cells (CHO-K1) and a strain of CHO cells which expresses the human Bcl-2 triangle gene (CHO-bcl-2). In this study, the following questions were asked: (1) Does an acute hydrodynamic force induce apoptosis in wild-type CHO and CHO-bcl-2 cells? (2) Does the type of culture media make a difference with respect to the induction of apoptosis or necrosis? and (3) Does culture history affect induction of apoptosis or necrosis? The results obtained with this new flow contraction device and corresponding computer simulations are consistent with previously published studies with respect to the level of energy dissipation rate (EDR) required to create significant cell lysis. Second, while detectable relative to the control in the T-flask experiments, only a small fraction of the cells become apoptotic when exposed to a sub-lysis level of EDR (<10(8) W x m(-3)). Third, cells cultured in suspension with serum free media do not exhibit any higher or lower sensitivity (with respect to apoptosis) to various levels of EDR when compared to control cultures grown in T-flask and serum containing media; on the other hand, necrosis is significantly increased in experiments performed on suspended cells without serum. Fourth, the addition of the Bcl-2 gene product might slightly reduce the occurrence of apoptosis in T-flask culture; however, the baseline response is so low that the difference is insignificant.
开发并建立了第二代流体收缩装置模型，该装置可使细胞受到明确的流体动力作用。利用该系统对野生型中国仓鼠卵巢细胞（CHO-K1）和表达人Bcl-2三角基因的CHO细胞株（CHO-bcl-2）进行了研究。在本研究中，提出了以下问题：（1）急性流体动力是否会诱导野生型CHO和CHO-bcl-2细胞凋亡？（2）培养基类型对凋亡或坏死的诱导是否有影响？以及（3）培养历史是否会影响凋亡或坏死的诱导？使用这种新型流体收缩装置获得的结果以及相应的计算机模拟结果，在产生显著细胞裂解所需的能量耗散率（EDR）水平方面，与先前发表的研究结果一致。其次，虽然在T型瓶实验中相对于对照组可检测到，但当暴露于亚裂解水平的EDR（<10^8 W·m^-3）时，只有一小部分细胞发生凋亡。第三，与在T型瓶中生长且含有血清的培养基中的对照培养物相比，在无血清培养基中悬浮培养的细胞对各种水平的EDR没有表现出更高或更低的（凋亡）敏感性；另一方面，在无血清的悬浮细胞实验中，坏死显著增加。第四，添加Bcl-2基因产物可能会略微降低T型瓶培养中凋亡的发生率；然而，基线反应非常低，以至于差异不显著。
