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Article

Exosome-Based Molecular Transfer Activity of Macrophage-Like Cells Involves Viability of Oral Carcinoma Cells: Size Exclusion Chromatography and Concentration Filter Method

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Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8525, Japan
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Department of Dental Anesthesiology and Special Care Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8525, Japan
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Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8525, Japan
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Department of Clinical Engineering, Faculty of Life Sciences, Hiroshima Institute of Technology, Hiroshima 731-5193, Japan
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Department of Biochemistry, Ain Shams University Faculty of Science, Cairo 11566, Egypt
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Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8502, Japan
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Research Program for Undergraduate Students, Okayama University Dental School, Okayama 700-8525, Japan
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O-NECUS Program of Okayama University Dental School, Department of Endodontics, School of Stomatology, China Medical University, Shenyang 110002, China
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Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan
*
Author to whom correspondence should be addressed.
Academic Editors: Carlos Salomon Gallo and Andreas Moller
Cells 2021, 10(6), 1328; https://doi.org/10.3390/cells10061328
Received: 19 April 2021 / Revised: 11 May 2021 / Accepted: 24 May 2021 / Published: 27 May 2021
Extracellular vesicles (EV) heterogeneity is a crucial issue in biology and medicine. In addition, tumor-associated macrophages are key components in cancer microenvironment and immunology. We developed a combination method of size exclusion chromatography and concentration filters (SEC-CF) and aimed to characterize different EV types by their size, cargo types, and functions. A human monocytic leukemia cell line THP-1 was differentiated to CD14-positive macrophage-like cells by stimulation with PMA (phorbol 12-myristate 13-acetate) but not M1 or M2 types. Using the SEC-CF method, the following five EV types were fractionated from the culture supernatant of macrophage-like cells: (i) rare large EVs (500–3000 nm) reminiscent of apoptosomes, (ii) EVs (100–500 nm) reminiscent of microvesicles (or microparticles), (iii) EVs (80–300 nm) containing CD9-positive large exosomes (EXO-L), (iv) EVs (20–200 nm) containing unidentified vesicles/particles, and (v) EVs (10–70 nm) containing CD63/HSP90-positive small exosomes (EXO-S) and particles. For a molecular transfer assay, we developed a THP-1-based stable cell line producing a GFP-fused palmitoylation signal (palmGFP) associated with the membrane. The THP1/palmGFP cells were differentiated into macrophages producing palmGFP-contained EVs. The macrophage/palmGFP-secreted EXO-S and EXO-L efficiently transferred the palmGFP to receiver human oral carcinoma cells (HSC-3/palmTomato), as compared to other EV types. In addition, the macrophage-secreted EXO-S and EXO-L significantly reduced the cell viability (ATP content) in oral carcinoma cells. Taken together, the SEC-CF method is useful for the purification of large and small exosomes with higher molecular transfer activities, enabling efficient molecular delivery to target cells. View Full-Text
Keywords: macrophage; exosomes; extracellular vesicles; molecular transfer; size exclusion chromatography and concentration filter (SEC-CF) method; heat shock proteins; oral carcinoma macrophage; exosomes; extracellular vesicles; molecular transfer; size exclusion chromatography and concentration filter (SEC-CF) method; heat shock proteins; oral carcinoma
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MDPI and ACS Style

Lu, Y.; Eguchi, T.; Sogawa, C.; Taha, E.A.; Tran, M.T.; Nara, T.; Wei, P.; Fukuoka, S.; Miyawaki, T.; Okamoto, K. Exosome-Based Molecular Transfer Activity of Macrophage-Like Cells Involves Viability of Oral Carcinoma Cells: Size Exclusion Chromatography and Concentration Filter Method. Cells 2021, 10, 1328. https://doi.org/10.3390/cells10061328

AMA Style

Lu Y, Eguchi T, Sogawa C, Taha EA, Tran MT, Nara T, Wei P, Fukuoka S, Miyawaki T, Okamoto K. Exosome-Based Molecular Transfer Activity of Macrophage-Like Cells Involves Viability of Oral Carcinoma Cells: Size Exclusion Chromatography and Concentration Filter Method. Cells. 2021; 10(6):1328. https://doi.org/10.3390/cells10061328

Chicago/Turabian Style

Lu, Yanyin, Takanori Eguchi, Chiharu Sogawa, Eman A. Taha, Manh T. Tran, Toshiki Nara, Penggong Wei, Shiro Fukuoka, Takuya Miyawaki, and Kuniaki Okamoto. 2021. "Exosome-Based Molecular Transfer Activity of Macrophage-Like Cells Involves Viability of Oral Carcinoma Cells: Size Exclusion Chromatography and Concentration Filter Method" Cells 10, no. 6: 1328. https://doi.org/10.3390/cells10061328

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