Authors: Duan Sujuan, Cui Wenwen, Liu Xing, Huang Guofu
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Objective
To explore the effects of shear stress on morphology, adhesion and proliferation of human umbilical cord blood mesenchymal stem cells(hUC-MSCs).
Methods
The hUC-MSCs were cultured in vitro until confluence and then placed in a flow system.The cells were subjected to different shear stress (1, 2, 3, 4 dye/cm2) for 2 hours and 6 hours, and no shear stress treatment cells served as a static control.The morphological changes of hUC-MSCs in different groups were analyzed by phase contrast microscopy and immunofluorescence.The mRNA expression levels of intercellular adhesion molecule-1 (ICAM-1) and Ki67 were detected by real-time PCR.
Results
Compared with the static control group, the hUC-MSCs cells were arranged in the direction of fluid after treated with shear stress.The immunofluorescence results showed that the cytoskeletal protein F-actin filaments was prolonged after shear stress.The cytoskeleton was further elongated in the 2 dye/cm2 for 6 hours group when compared with 2 dye/cm2 for 2 hours group, and the cytoskeleton was loosened when time extended to 6 hours.Real-time PCR results showed that the relative expressions of ICAM-1 mRNA and Ki67 mRNA in the static control group and different gradient shear stress groups were significantly different, with significant differences among them (F=17.141, P=0.000; F=11.336, P=0.001). The relative expression of ICAM-1 mRNA in the 1, 2, 3, 4 dye/cm2 shear stress group was 2.74±0.32, 9.77±1.19, 6.70±0.92 and 5.69±0.72, respectively, which was significantly higher than 1.00±0.28 in the static control group, with significant differences between them (all at P<0.05). The relative expression of Ki67 mRNA in the 3 dye/cm2 and 4 dye/cm2 shear stress group was 0.39±0.09 and 0.04±0.02, respectively, which was significantly lower than 1.00±0.24 in the static control group, with statistically significant differences between them (both at P<0.05).
Conclusions
After treated with fluid shear stress, hUC-MSCs are arranged in the direction of fluid.Shear stress can promote the adhesion of hUC-MSCs.As the increase of shear stress intensity, cell proliferation is inhibited.