Three-Dimensional Culture of Umbilical Cord Mesenchymal Stem Cells Effectively Promotes Platelet Recovery in Immune Thrombocytopenia

Abstract

Mesenchymal stem cells (MSCs) can effectively regulate immune cell functions and therefore are promising for the treatment of autoimmune disorders, such as immune thrombocytopenia (ITP). Recent research has shown that three-dimensional (3D) culture method have many advantages over conventional culture with respect to MSC secretion and immunogenicity. In this study, 2D and 3D cultured MSCs were used to evaluate cytokine secretion, extracellular matrix (ECM) gene expression, immune regulatory activity, and therapeutic effects in a mouse model of ITP. MSCs cultured on scaffolds had higher expression levels of immune regulatory genes, such as IDO1, HLA-G, and PTGS2, and greater inhibitory activity against lymphocyte activation that those of 2D-MSCs. In addition, 3D-MSCs exhibited higher ECM expression and greater protection against interferon-γ (IFN-γ)-induced apoptosis. In a mouse study, ITP was induced by guinea pig anti-mouse platelet serum injections. Based on enzyme-linked immunosorbent assays, serum levels of the suppressive cytokine IL-10 were higher and IFN-γ levels were lower after intravenous injection with 3D-MSCs and with 2D-MSCs. Additionally, 3D-MSCs improved the body weight, spleen index, and platelet index relative to those for 2D-MSCs. Bone marrow homing was also significantly enhanced in the 3D group. Therefore, the 3D culture of MSCs is an effective technique for the treatment of ITP.

The Immunoregulatory Effect of Mechanical Stress on Three-dimensional Cultured Mesenchymal Stem Cells After Extensive Expansion

“…One research also indicated that the expressions of HLA-G, IDO1, PTGS2 and TGF-β1, which were closely related to immune regulation, were higher in 3D culture than those in 2D culture. Further, 3D cultured MSCs could inhibit the proliferation of activated lymphocytes more effectively [17]. Here, we compared the stemness and immunomodulatory ability between 2D and 3D UC-MSCs and found that 3D culture did preserve the stemness, T cell proliferative inhibition and the expression of anti-in ammatory cytokines such as IL-10 and TGF-β of long-passaged UC-MSCs.…”

“…One study found several genes closely related to immune regulation, such as human leukocyte antigen (HLA)-G, indoleamine 2,3-dioxygenase (IDO)-1, prostaglandin synthase (PTGS)-2 and transforming growth factor (TGF)-β1 were more highly expressed after 3D culture than that after 2D culture. Further, 3D cultured MSCs could inhibit the proliferation of activated lymphocytes more effectively [17]. However, the mechanism of maintaining cellular immunomodulatory ability in 3D culture has not been reported, and exploring this mechanism provides an experimental basis for further elucidating the immunomodulatory characteristic of MSCs in 3D culture.…”

The Mechanistic Effects and Clinical Applications of Various Derived Mesenchymal Stem Cells in Immune Thrombocytopenia

“…However, the development of 3D culture methods has improved the ability to amplify BM-MSCs. The BM-MSCs amplified through this method are superior to those produced by traditional culture methods in terms of secretion and immunogenicity, and they also have been found to improve the body weight, spleen index, platelet index, and bone marrow homing ability in a mouse model of ITP [54]. Cytological experiments performed in vitro have also shown that BM-MSCs regulate the balance of Th1/Th2 cell activity in ITP, thereby inhibiting the secretion of IL-2 and IFN-γ by Th1 cells and promoting the secretion of IL-4 by Th2 cells, which then upregulate CD4+ CD25+ CD25+ T cells and induce immune tolerance [55].…”

The Extensive Regulation of MicroRNA in Immune Thrombocytopenia

“… 156 , 157 Due to the immunomodulatory effect of MSCs, MSCs transplantation has shown a certain therapeutic effect on ITP in vivo and in vitro, and in the clinic. 158 – 161 …”