Xeno-Free Condition Enhances Therapeutic Functions of Human Wharton's Jelly-Derived Mesenchymal Stem Cells against Experimental Colitis by Upregulated Indoleamine 2,3-Dioxygenase Activity.

Ji Yeon Kang,Hae-Ri Lee,Mi-Kyung Oh,Hansol Joo,Hyun Sung Park,Kyung-Rok Yu,Jieun Kim,Il-Hoan Oh,Dong-Hoon Chae

doi:10.3390/jcm9092913

Abstract

The therapeutic applications of mesenchymal stem cells (MSCs) have been actively explored due to their broad anti-inflammatory and immunomodulatory properties. However, the use of xenogeneic components, including fetal bovine serum (FBS), in the expansion media might pose a risk of xenoimmunization and zoonotic transmission to post-transplanted patients. Here, we extensively compared the physiological functions of human Wharton’s jelly-derived MSCs (WJ-MSCs) in a xeno-free medium (XF-MSCs) and a medium containing 10% FBS (10%-MSCs). Both groups showed similar proliferation potential; however, the 10%-MSCs showed prolonged expression of CD146, with higher colony-forming unit-fibroblast (CFU-F) ability than the XF-MSCs. The XF-MSCs showed enhanced adipogenic differentiation potential and sufficient hematopoietic stem cell (HSC) niche activity, with elevated niche-related markers including CXCL12. Furthermore, we demonstrated that the XF-MSCs had a significantly higher suppressive effect on human peripheral blood-derived T cell proliferation, Th1 and Th17 differentiation, as well as naïve macrophage polarization toward an M1 phenotype. Among the anti-inflammatory molecules, the production of indoleamine 2,3-dioxygenase (IDO) and nitric oxide synthase 2 (NOS2) was profoundly increased, whereas cyclooxygenase-2 (COX-2) was decreased in the XF-MSCs. Finally, the XF-MSCs had an enhanced therapeutic effect against mouse experimental colitis. These findings indicate that xeno-free culture conditions improved the immunomodulatory properties of WJ-MSCs and ex vivo-expanded XF-MSCs might be an effective strategy for preventing the progression of colitis.

Highlights

Human mesenchymal stem cells (MSCs) have been isolated and expanded from many different adult and embryonic tissues [1]
When the surface phenotype was compared, the characteristic Wharton’s jelly-derived MSCs (WJ-MSCs) immune phenotypes of CD29, CD44, CD73, CD90, and CD105 were expressed in XF and 10% fetal bovine serum (FBS) conditions, but the expression of CD146 was reduced in long-term expanded cells under XF conditions (Figure 1C and Supplementary Figure S1)
To confirm the colony-forming unit-fibroblast (CFU-F) ability of 10%-FBS and XF-MSCs, the cells were plated at low density in a 100-mm dish

Summary

Introduction

Human mesenchymal stem cells (MSCs) have been isolated and expanded from many different adult and embryonic tissues [1]. The immunomodulatory properties of MSCs involve soluble factors and cell contact-dependent processes in response to an inflammatory environment and immune cells [4]. MSC-mediated immunomodulation regulates both the innate and the adaptive immune system by inhibiting the proliferation of CD4 and CD8 T cells [5] and pro-inflammatory type 1 helper T cells (Th1) as well as interferon-gamma (IFN-γ) production by Th1 cells [6,7]. Tumor necrosis factor (TNF)-α-mediated activation of MSCs promotes the polarization of naïve macrophages toward anti-inflammatory type (M2) macrophages and inhibits differentiation into a pro-inflammatory type phenotype (M1) and dendritic cells (DCs) [8,9]. We and others have shown that the soluble factors proposed to be involved during the immunomodulatory process include prostaglandin E2 (PGE2), indoleamine 2,3-dioxygenase (IDO), nitric oxide, transforming growth factor-beta (TGF-β), IL-6, and IL-10 [10,11]

Methods

Results

Conclusion