Immunosuppressive mesenchymal stem cells aggregates incorporating hydrogel microspheres promote an in vitro invasion of cancer cells

Teruki Nii; Yasuhiko Tabata

doi:10.1016/j.reth.2021.11.006

Immunosuppressive mesenchymal stem cells aggregates incorporating hydrogel microspheres promote an in vitro invasion of cancer cells

Regen Ther. 2021 Dec 10:18:516-522. doi: 10.1016/j.reth.2021.11.006. eCollection 2021 Dec.

Authors

Teruki Nii¹, Yasuhiko Tabata¹

Affiliation

¹ Laboratory of Biomaterials, Institute for Frontier Life and Medical Sciences, Kyoto University, 53 Kawara-cho Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.

Abstract

Introduction: The objective of this study is to design a co-culture system of cancer cells and three-dimensional (3D) mesenchymal stem cells (MSC) aggregates for the in vitro evaluation of cancer invasion.

Methods: First, the MSC of an immunosuppressive phenotype (MSC2) were prepared by the MSC stimulation of polyriboinosinic polyribocytidylic acid. By simple mixing MSC2 and gelatin hydrogel microspheres (GM) in a U-bottomed well of 96 well plates which had been pre-coated with poly (vinyl alcohol), 3D MSC2 aggregates incorporating GM were obtained. The amount of chemokine (C-C motif) ligand 5 (CCL5) secreted from the MSC2 aggregates incorporating GM. Finally, an invasion assay was performed to evaluate the cancer invasion rate by co-cultured cancer cells and the 3D MSC2 incorporating GM.

Results: The amount of CCL5 secreted for the 3D MSC2 aggregates incorporating GM was significantly higher than that of two-dimensional (2D) MSC, 2D MSC2, and 3D MSC aggregates incorporating GM. When MDA-MB-231 human breast cancer cells were co-cultured with the 3D MSC2 aggregates incorporating GM, the invasion rate of cancer cells was significantly high compared with that of 2D MSC or 2D MSC2 and 3D MSC aggregates incorporating GM. In addition, high secretion of matrix metalloproteinase-2 was observed for the 3D MSC2 aggregates/cancer cells system.

Conclusions: It is concluded that the co-culture system of 3D MSC2 aggregates incorporating GM and cancer cells is promising to evaluate the invasion of cancer cells in vitro.

Keywords: (CCL)5, chemokine (C–C motif) ligand; 2D, two-dimensional; 3D, three-dimensional; Anti-cancer drug screening; CAF, cancer-associated fibroblasts; Cancer invasion model; DDW, double-distilled water; DMEM, Dulbecco's modified Eagle's medium; ELISA, enzyme-linked immunosolvent assay; FCS, fetal calf serum; GM, gelatin hydrogel microspheres; Gelatin hydrogel microspheres; MEM, minimum essential medium; MMP, matrix metalloproteinase; MSC, mesenchymal stem cells; MSC2, MSC of an immunosuppressive phenotype; Mesenchymal stem cells; PBS, phosphate buffered-saline; PVA, poly (vinyl alcohol); TAM, tumor-associated macrophages; Three-dimensional cell culture.