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《Oncology Progress》 2018-02
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Effect of substrate stiffness on cell growth of HeLa cells in vitro

ZHAO Xuanyu;JIAO Simeng;SONG Dan;CHEN Jiao;SHANG Ruotian;ZHANG Qiang;LIU Sidi;KONG Weimin;HAN Dong;Department of Gynecologic Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University;National Center for Nanoscience and Technology;  
Objective To explore the effect of substrate stiffness on the growth of cervical cancer HeLa cells in vitro. Method Human cervical cancer cell line HeLa was cultured in hydrogel culture dishes with the substrate stiffness of 0.5, 5.0 and 25.0 kPa, as well as in ordinary plastic/glass culture dishes with the substrate stiffness of 1.0×106 kPa under same conditions. Cell morphological structure was observed using inverted microscope and the environmental scanning electron microscope; cell nucleus and cytoskeleton were observed under the single-photon laser-scanning confocal microscope. Cell proliferation activity was detected using the cell counting kit-8 and survival curves were plotted. Result As the substrate stiffness increased, HeLa cells tended to spread, and adhered to culture dishes more firmly, with polygon or spindle morphology; The maximum/minimum diameter and cell spread area among dishes with different stiffness showed statistically significant difference(F=15.54, 77.21, P0.01); As the substrate stiffness increased, higher stiffness of dishes promoted cells to grow more long pseudopodia and expressed increased cytoskeletal stress fibers; the culture time of 48 h was used to calculate the time for cell doubling, and it was found that, the doubling time of HeLa cells cultured in dishes with substrate stiffness of 0.5, 5.0, 25.0, and 1.0×106 kPa was 60.40, 39.43, 27.79 and 26.34 h, respectively, suggesting that the cell doubling time decreased, while cell proliferation accelerated as the substrate stiffness increased. Conclusion Higher substrate stiffness is favorable for human cervical cancer cells HeLa to spread, and may promote cytoskeletal distribution and cell proliferation.
【Fund】: 北京市科学技术委员会生命科学领域前沿技术培育课题(Z161100000116030)
【CateGory Index】: R737.33
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