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. 2020 Jul 20;9(1):1795364. doi: 10.1080/20013078.2020.1795364

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© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Characterization and Evaluation of itTBC-EVs and a scheme for induction of immune-tolerized stem cells (itSCs) and immune-tolerized EVs (itSC-EVs) through cultivation on HA-Matrix^Decidua containing itTBC-EVs with temperature/vibration conditions. (a) NTA analysis of EVs derived from BeWo^Control cultured on a general culture plate without temperature/vibration conditions. (b) NTA analysis of EVs derived from BeWo^Matrix+T/V cultured on the HA-Matrix^Decidua with the in vivo-like temperature change and vibration conditions. BeWo^Matrix+T/V-EVs have more EV particles than BeWo^Control due to high proliferation rate during invasion to the HA-Matrix^Decidua. (c) The images of BeWo^Matrix+T/V-EVs using SEM (left) and TEM (right), respectively. (d)The expression of exosomal markers, CD9, CD63, and CD81, on BeWo^Matrix+T/V-EVs using ExoView. (e) The expression of HLA-G isoforms in BeWo^Matrix+T/V-EVs and BeWo^Control-EVs using ExoView. The expressions of 4H84, bound to α1 domain of HLA-G heavy chain (with blue bar), and MEM-G/9, bound to β2 m-associated HLA-G isoforms (with red bar) are much higher in BeWo^Matrix+T/V-EVs than BeWo^Control-EVs. (F) The images of HLA-G-bound EV spots (blue and red) captured with CD9 antibody using ExoView. (g) A diagram for two methods to induce immune-tolerized stem cells (itSCs) and immune-tolerized EVs (itSC-EVs), by co-cultivation with itTBCs (right) and by using itTBC-EVs (left) under the Ex-vivo culture system established herein.