Effect of peripheral blood-derived mesenchymal stem cells on macrophage polarization and Th17/Treg balance in vitro

Rui Yang; Hongfei Gao; Long Chen; Ning Fang; Hui Chen; Gongyu Song; Limei Yu; Qian Zhang; Tao Zhang

doi:10.1016/j.reth.2020.03.008

Effect of peripheral blood-derived mesenchymal stem cells on macrophage polarization and Th17/Treg balance in vitro

Regen Ther. 2020 May 16:14:275-283. doi: 10.1016/j.reth.2020.03.008. eCollection 2020 Jun.

Authors

Rui Yang¹, Hongfei Gao¹, Long Chen², Ning Fang¹, Hui Chen¹, Gongyu Song³, Limei Yu¹, Qian Zhang³, Tao Zhang¹

Affiliations

¹ Key Laboratory of Cell Engineering of Guizhou Province and Regenerative Medicine Centre, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou, China.
² Experimental Medical Centre, Dongfeng Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
³ Department of Human Anatomy, Zunyi Medical University, Zunyi, Guizhou, China.

Abstract

Introduction: Mesenchymal stem cells (MSCs) have always been the center of the experimental exploration of regenerative therapy together with other stem cells. Among with, peripheral blood-derived mesenchymal stem cells (PBMSCs) have been regarded as promising in clinical applications for its convenience of acquisition from peripheral blood. However, few reported experiments so far to elucidate the exact mechanisms of how PBMSC influence regeneration. As the ability of immunomodulatory is one of the crucial features that influence MSC to reconstruct impaired tissue, we decided to focus on the immunomodulatory abilities of PBMSCs and conducted experiments associated with macrophages and T lymphocytes, which are two main cell types that dominate the innate and acquired immunity. Therefore, a basis can be made from these experiments for applications of PBMSCs in regenerative therapy in the future.

Methods: A Transwell system was used for the coculturing of PBMSCs with macrophages. T lymphocytes were cultured directly with PBMSCs. Flow cytometry and immunochemistry were conducted for identifying the phenotypes. Immunomagnetic microspheres, ELISA and RT-qPCR were used to detect the expressions of relevant molecules or mRNAs.

Results: After coculturing PBMSCs with M0, the anti-inflammatory IL-10 was increased whereas the proinflammatory TNF-α decreased; the expression of CD11b, CD68, CD206, Arg-1, IL-10 and CCL-22 was up-regulated whereas IL-1β down-regulated. The expression of TGF-β, RORγt, Foxp3 and IL-10 was increased in the cocultured lymphocytes whereas IL-17 and IL-6 decreased; the ratio of CD4⁺IL-17⁺ Th17/CD25⁺Foxp3⁺ Treg was reduced.

Conclusion: The findings demonstrated that PBMSCs promoted the anti-inflammatory features of macrophages and the Th17/Treg system. PBMSCs are able to inhibit inflammation associated with these two immune cell systems, and thus provide insight into how PBMSCs achieve their immunomodulatory ability.

Keywords: Inflammation; Macrophages; Mesenchymal stem cells; Stem cells; T lymphocytes.