本研究設計圓錐平板式生物反應器，利用圓錐旋轉時帶動培養液產生剪應力，對平板上的細胞施予可控制的定量剪應力刺激值。使用此生物反應器，對大鼠骨髓幹細胞進行1天14小時的培養，並對其分別施加1.4、5.5、11、16.4 dyn/cm2的剪應力刺激。結果發現施予剪應力刺激的細胞其生長速度受到抑制，而刺激強度超過1.4 dyn/cm2後，細胞型態會隨著刺激強度增加而逐漸變差，細胞存活數目減少。此外，在1.4 dyn/cm2的4小時刺激強度下，將培養時間延長至3天又14小時，亦發現細胞生長速度受到抑制，且於細胞型態上，出現細胞開始拉長並有方向性排列的情況。 實驗結果顯示，本研究設計的圓錐平板裝置可穩定提供剪應力機械刺激，且能於體外穩定培養細胞。未來，對於經過剪應力刺激已出現形態變化的細胞，可於生化方面進行相關檢測，並給予細胞不同強度或週期性的剪應力刺激，觀察細胞增殖數量的生長變化。 We designed a cone-plate bioreactor. It produces fluid shear stress by rotating the cone that drives the culture medium to flow in a circular way. Rat bone marrow stem cells were cultured on coverslips placed on the plate. The cells experienced the shear stress of 1.4、5.5、11、16.4 dyn/cm2 respectively for 4 hours in 1 day and 14 hours in the bioreactor. Results showed the shear stresses changed the morphology and inhibited the cell growth when the shear magnitudes were over 1.4 dyn/cm2. We also found that, by applying shear stress of 1.4 dyn/cm2 for 4 hours within 3 days and 14 hours, the cells became elongated in shape and aligned to each other. The study demonstrates that the cone-plate bioreactor can provide well-defined stimulus to the cells cultured in vitro. The growth inhibition and shape changes of the cells by shears shall merit further research efforts to classify the underlying mechanisms and biochemical pathways.
