Comparison of ultracentrifugation and density gradient separation methods for isolating Tca8113 human tongue cancer cell line‑derived exosomes

Zhang,Zhuoyuan; Wang,Chenxing; Li,Tang; Liu,Zhe; Li,Longjiang

doi:10.3892/ol.2014.2373

Comparison of ultracentrifugation and density gradient separation methods for isolating Tca8113 human tongue cancer cell line‑derived exosomes

Authors:
- Zhuoyuan Zhang
- Chenxing Wang
- Tang Li
- Zhe Liu
- Longjiang Li
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Affiliations: State Key Laboratory of Oral Diseases, West China School of Stomatology, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: July 23, 2014 https://doi.org/10.3892/ol.2014.2373
Pages: 1701-1706

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Abstract

The aim of the present study was to compare the method of ultracentrifugation and density gradient separation for isolating Tca8113 human tongue squamous cell carcinoma cell line‑derived exosomes. The exosomes were obtained from the culture supernatant of cultured Tca8113 cells, respectively, followed by identification with transmission electron microscopy observation and western blot analysis. The two different methods were then compared by the morphology, the distribution range of the particle size and the concentration of proteins of the extracted exosomes. In vitro, Tca8113 cells can secrete a large amount of vesicle‑like structures, which are identified as exosomes by the presence of the surface markers, Hsp‑70 and Alix. The protein profile of the two products are almost the same, however the particle size distribution of the exosomes extracted with density gradient centrifugation are more limited, between 40‑120 nm, and these have a higher protein concentration. The results indicate that Tca8113 cells can secrete exosomes in vitro, and the density gradient separation methods for purifying exosomes is improved, which is helpful for future research and application of exosomes.

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