Comparison of angiogenic, cytoprotective, and immunosuppressive properties of human amnion- and chorion-derived mesenchymal stem cells

Abstract

Although mesenchymal stem cells (MSCs) can be obtained from the fetal membrane (FM), little information is available regarding biological differences in MSCs derived from different layers of the FM or their therapeutic potential. Isolated MSCs from both amnion and chorion layers of FM showed similar morphological appearance, multipotency, and cell-surface antigen expression. Conditioned media obtained from amnion- and chorion-derived MSCs inhibited cell death caused by serum starvation or hypoxia in endothelial cells and cardiomyocytes. Amnion and chorion MSCs secreted significant amounts of angiogenic factors including HGF, IGF-1, VEGF, and bFGF, although differences in the cellular expression profile of these soluble factors were observed. Transplantation of human amnion or chorion MSCs significantly increased blood flow and capillary density in a murine hindlimb ischemia model. In addition, compared to human chorion MSCs, human amnion MSCs markedly reduced T-lymphocyte proliferation with the enhanced secretion of PGE2, and improved the pathological situation of a mouse model of acute graft-versus-host disease. Our results highlight that human amnion- and chorion-derived MSCs, which showed differences in their soluble factor secretion and angiogenic/immuno-suppressive function, could be ideal cell sources for regenerative medicine.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Characterization of human amnion- and chorion-derived MSCs.

(A) Representative photographs of human amnion and chorion. (B) Photographs of cultured MSCs obtained from human amnion and chorion at passage 3. Scale bars = 500 µm. (C) Relative cell number of amnion- and chorion-derived MSCs at each passage. (D) FACS analysis of amnion and chorion MSCs. (E, F) Differentiation of amnion and chorion MSCs into adipocytes (E) and osteocytes (F). Scale bars = 100 (E) and 50 (F) µm.

Figure 2. Growth factor secretion and the cytoprotective effect of amnion and chorion MSCs.

(A–D) Cytoprotective effect of FM MSC-derived conditioned medium was analyzed by the MTS assay (A, B) and caspase-3 activity (C, D) in HUVECs (A, C) and cardiomyocytes (B, D). Values are mean ± SEM. *p

Figure 3. Angiogenic potential of amnion and chorion MSCs against hindlimb ischemia.

(A) Representative images of serial hindlimb blood perfusion. Blood perfusion of ischemic hindlimb increased in the amnion and chorion MSC groups at day 5. (B) Quantitative analysis of hindlimb blood perfusion with the LDPI index, the ratio of ischemic to non-ischemic hindlimb blood perfusion. (C) Representative photographs of immunohistochemistry with anti-CD31 antibody. Scale bars = 100 µm. (D) Quantitative analysis of capillary density in ischemic hindlimb muscle at day 5 among the control, amnion, and chorion MSC groups. Capillary density is shown as the capillary-to-muscle-fiber ratio. Data are mean ± SEM. **p

Figure 4. Immunosuppressive property of amnion and chorion MSCs.

(A) Inhibition of human CD4+ T cell proliferation upon co-culture with human amnion, chorion, and bone marrow MSCs. (B) The concentration of PGE2 in FM-MSC-conditioned medium was measured by ELISA. Amnion MSCs secreted a significant amount of PGE2 compared with chorion MSCs. (C, D) Effect of human amnion (C) or chorion (D) MSC transplantation in a murine GVHD model. Treatment with amnion MSCs significantly reduced recipient weight loss in a mouse model of GVHD. *p