Table 1.
Primers used in this study.
Fig 1.
Morphology and Growth Kinetics of hfC-MSCs compared to hBM-MSCs.
Representative photomicrographs (10X, 20μm) of (A) Human fetal cardiac mesenchymal stem cells (hfC-MSCs); (B) Bone marrow mesenchymal stem cells (hBM-MSCs) showing spindle shaped morphology at 5th passage; (C) Growth kinetics of hfC-MSCs and hBM-MSCs seeded at a density of 1,000 cells per cm2.
Fig 2.
Phenotypic characteristics of hfC-MSCs compared to BM-MSCs.
Representative bar graphs showing a comparison of the expression of CD73, CD90, CD105, SSEA-4, CD117, CD34, CD45, HLA-DR and CD31 on hfC-MSCs and hBM-MSCs as demonstrated by flow cytometry. Values are mean ± SE of three independent experiments of both the cell types at passage-5.
Fig 3.
Expression of pluripotency and embryonic markers and cardiovascular genes by hfC-MSCs and hBM-MSCs.
Representative photomicrographs (40X, 20μm) of human fetal cardiac mesenchymal stem cells (hfC-MSCs) showing expression of OCT-4 (A: OCT-4; B: hoechst dye), Nanog (C: Nanog; D: hoechst dye), SOX-2 (E: SOX-2; F: hoechst dye) and representative photomicrographs of (40X, 20μm) of human bone marrow mesenchymal stem cells (hfC-MSCs) showing expression of OCT-4 (G: OCT-4; H: hoechst dye), Nanog (I: Nanog; J: hoechst dye), SOX-2 (K: SOX-2; L: hoechst dye).
Fig 4.
(A-E) Bar graphs showing relative gene expression of GATA-4, Isl-1, Flk-1, NKX2.5 and MDR-1 transcripts in human fetal cardiac mesenchymal stem cells (hfC-MSCs) compared to human bone marrow mesenchymal stem cells (hBM-MSCs) in 5th passage (P5) as demonstrated by real time PCR. Data shown are mean ± SE of 3 experiments. *p<0.05 of hBM-MSCs vs human fC-MSCs.
Fig 5.
Differentiation of hfC-MSCs into cardiovascular cells.
Representative immunocytochemistry images (40X, 20μm) showing differentiation of human fetal cardiac stem cells (hfC-MSCs) into Cardiomyocytes (B: Troponin-T (cTnT); Endothelial cells (D: CD31); Smooth Muscle Cells (F: Smooth Muscle- myosin heavy chain (SM-MHC); (A, C and E, were control cells without induction medium showing only hoechst dye). Data shown are from three independent experiments at passage 3–5.
Fig 6.
hfC-MSCs inhibit PHA-induced proliferation of lymphocytes.
(A) PBMCs (1 × 105 cells) stimulated with or without PHA (5 μg/ mL) in the presence or absence of irradiated hfC-MSCs (1 × 104–5 × 104 cells). Data are expressed as the mean ± SE of three independent experiments. *p<0.05 (Control PBMCs vs. PBMCs+ MSCs), NS: Not significant. (B) TGF-β levels were analyzed in the culture supernatants of co-cultured hfC-MSCs and PBMCs stimulated with or without PHA. PBMCs (1 × 105 cells) cultured with PHA (5 μg/mL) in the presence or absence of hfC-MSCs (1 × 104–5× 104 cells). Data are expressed as the mean ± SE of three independent experiments. *p<0.05 (Control PBMC vs. PBMC+ MSC), NS: Not significant. (C) IL-10 levels were analysed in the culture supernatants of co-cultured hfC-MSCs and PBMCs stimulated with or without PHA. PBMCs (1 × 105 cells) cultured with PHA (5 μg/mL) in the presence or absence of hfC-MSCs (1 × 104–5× 104 cells). Data are expressed as mean ± SE of three independent experiments. *p<0.05 (Control PBMCs vs. PBMCs+ MSCs), NS: Not significant.