Extracellular Vesicles Derived From Canine Mesenchymal Stromal Cells in Serum Free Culture Medium Have Anti-inflammatory Effect on Microglial Cells

Yukina Kuwahara; Karin Yoshizaki; Hidetaka Nishida; Hiroaki Kamishina; Sadatoshi Maeda; Katsura Takano; Naoki Fujita; Ryohei Nishimura; Jun-Ichiro Jo; Yasuhiko Tabata; Hideo Akiyoshi

doi:10.3389/fvets.2021.633426

Extracellular Vesicles Derived From Canine Mesenchymal Stromal Cells in Serum Free Culture Medium Have Anti-inflammatory Effect on Microglial Cells

Front Vet Sci. 2021 Apr 28:8:633426. doi: 10.3389/fvets.2021.633426. eCollection 2021.

Authors

Affiliations

¹ Joint Department of Veterinary Medicine, The United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan.
² Department of Veterinary Surgery, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Japan.
³ Department of Integrative Physiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Japan.
⁴ Department of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
⁵ Laboratory of Biomaterials, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.

Abstract

Mesenchymal stem/stromal cells (MSCs) have been used as cell sources for treating dogs with naturally-occurring diseases. Extracellular vesicles (EVs) derived from MSCs are now recognized as pivotal to modulating the immune response and supporting tissue repair. Manufacture of MSC-EVs for clinical application mandates removal of the xeno-proteins, including fetal bovine serum. The objective of this study was to examine whether canine MSCs survived and secreted EVs in serum-free medium (SFM) conditions and to assess the immunomodulatory effect of EVs in vitro. Canine MSCs were found to survive and secrete EVs under SFM conditions. The surface markers of MSCs in the SFM were similar to MSCs in complete culture medium. Canine MSC-EVs had a diameter of ~300 nm and were positive for EV markers. MSC-derived EVs from the serum-free condition reduced the levels of IL-1β by BV-2 cells in response to LPS stimulation. These results warrant further studies of the use of SFM for producing EVs derived from canine MSCs.

Keywords: anti-inflammatory; canine; exosome; extracellular vesicles; mesenchymal stromal cells; serum free culture.