He Q, Cahill C J, Spiro M J
Department of Medicine, Harvard Medical School, Boston, MA, USA.
J Mol Cell Cardiol. 1996 May;28(5):1177-86. doi: 10.1006/jmcc.1996.0108.
Cardiac myocytes isolated from adult rat ventricles have been maintained in a stable, differentiated state for prolonged periods by the use of suspension culture on hydrophobic tissue culture inserts or agarose-coated plates. The success of this procedure depends on the use of low-serum media to prevent myocyte-myocyte interaction and proliferation of any residual endothelial cells. Myocytes cultured in this manner retain many of their structural characteristics, suggesting that maintenance of their elongated irregular shape is not dependent on interaction with extracellular matrix. They also exclude trypan blue, can be vitally stained by the uptake and reduction of the tetrazolium dye MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide], synthesize myosin and when returned to adhesive surfaces are capable of attachment and attendant dedifferentiation. Stability of the myocytes in suspension permits their use in co-culture experiments; specifically, myocytes separated from endothelial cells by the hydrophobic membrane of the tissue culture insert stimulated proliferation of the latter cells, suggesting this to be a useful system for studying myocyte-endothelial cell interaction.
从成年大鼠心室分离出的心肌细胞，通过在疏水组织培养插入物或琼脂糖包被的平板上进行悬浮培养，已能长时间维持在稳定的分化状态。该方法的成功取决于使用低血清培养基，以防止心肌细胞间的相互作用以及任何残留内皮细胞的增殖。以这种方式培养的心肌细胞保留了许多结构特征，这表明维持其细长不规则形状并不依赖于与细胞外基质的相互作用。它们还能排斥台盼蓝，可通过摄取和还原四氮唑染料MTT [3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑] 进行活体染色，合成肌球蛋白，并且当放回黏附表面时能够附着并随之发生去分化。心肌细胞在悬浮状态下的稳定性使其可用于共培养实验；具体而言，通过组织培养插入物的疏水膜与内皮细胞分离的心肌细胞刺激了后者细胞的增殖，这表明这是一个研究心肌细胞-内皮细胞相互作用的有用系统。
