Journal: Journal of Animal Reproduction and Biotechnology

Article Title: Limited in vitro differentiation of porcine induced pluripotent stem cells into endothelial cells

doi: 10.12750/jarb.38.3.109

Figure Lengend Snippet: Fig. 3. Immunocytochemistry of pluripotency-associated markers in differentiated cells derived from piPSCs cultured in differentiation media. Immunostaining images of pluripotency-associated markers (OCT-3/4, SOX2 and NANOG) in porcine differentiated cells derived from piPSCs (OSKMNL) cultured in four differentiation media [StemDiff APEL TM 2 medium (APEL), StemDiff APEL TM 2 medium supple- mented with 50 ng/mL of Vascular Endothelial Growth Factor (APEL + VEGF), Endothelial Cell Basal Medium-2 (EBM-2) or Endothelial Cell Basal Medium-2 supplemented with 50 ng/mL of Vascular Endothelial Growth Factor (EBM-2 + VEGF)] on cultured plates coated with matrigel ® (1:40 dilution with DMEM/F-12 medium) for 8 days. Blue: Staining of Hoechst 33342, Red: Staining of OCT-3/4, SOX2 and NANOG protein. Merge: Hoechst 33342 signal and OCT-3/4, SOX2 and NANOG protein. Scale bar = 50 µm.

Article Snippet: In vitro differentiation of piPSCs into endothelial cells For in vitro differentiation of piPSCs into endothelial cells, piPSCs (OSKMNL) were passaged by manually picking the colonies and then seeded onto 0.5% gelatin-coated culture plates in four differentiation media [Endothelial Cell Basal Medium 2 (EBM-2; Lonza, Muenchensteinerstrasse, Basel, Switzerland), Endothelial Cell Basal Medium 2 supplemented with 50 ng/mL of Vascular Endothelial Growth Factor (EBM-2 + VEGF; R&D Systems, Minneapolis, MN, USA), StemDiff APELTM 2 medium (APEL-2; STEMCELL Technologies, Vancouver, BC, Canada), StemDiff APELTM 2 medium supplemented with 50 ng/mL of Vascular Endo- thelial Growth Factor (APEL-2 + VEGF)] on cultured plates coated with matrigel® (CORNING, NY, USA) (1:40 dilution with DMEM/F-12 medium) for 8 days at 39°C.

Techniques: Immunocytochemistry, Derivative Assay, Cell Culture, Immunostaining, Staining

Journal: Journal of Animal Reproduction and Biotechnology

Article Title: Limited in vitro differentiation of porcine induced pluripotent stem cells into endothelial cells

doi: 10.12750/jarb.38.3.109

Figure Lengend Snippet: Fig. 5. Flow cytometry of endothelial cell marker (CD-31) in differentiated cells derived from piPSCs cultured in differentiation media. Flow cytometry of endothelial cell marker (CD-31) expression in porcine differentiated cells derived from piPSCs (OSKMNL) cultured in four differentiation media [StemDiff APEL TM 2 medium (APEL), StemDiff APEL TM 2 medium supplemented with 50 ng/mL of Vascular En- dothelial Growth Factor (APEL + VEGF), Endothelial Cell Basal Medium-2 (EBM-2) and Endothelial Cell Basal Medium-2 supplemented with 50 ng/mL of Vascular Endothelial Growth Factor (EBM-2 + VEGF)] on cultured plates coated with matrigel ® (1:40 dilution with DMEM/F-12 medium) for 8 days was analyzed. Swine umbilical cord vein endothelial cells (SUVEC) were used as the positive control. Cells in blue were treated with PE-conjugated CD-31 and cells in red were not treated with PE-conjugated CD-31.

Article Snippet: In vitro differentiation of piPSCs into endothelial cells For in vitro differentiation of piPSCs into endothelial cells, piPSCs (OSKMNL) were passaged by manually picking the colonies and then seeded onto 0.5% gelatin-coated culture plates in four differentiation media [Endothelial Cell Basal Medium 2 (EBM-2; Lonza, Muenchensteinerstrasse, Basel, Switzerland), Endothelial Cell Basal Medium 2 supplemented with 50 ng/mL of Vascular Endothelial Growth Factor (EBM-2 + VEGF; R&D Systems, Minneapolis, MN, USA), StemDiff APELTM 2 medium (APEL-2; STEMCELL Technologies, Vancouver, BC, Canada), StemDiff APELTM 2 medium supplemented with 50 ng/mL of Vascular Endo- thelial Growth Factor (APEL-2 + VEGF)] on cultured plates coated with matrigel® (CORNING, NY, USA) (1:40 dilution with DMEM/F-12 medium) for 8 days at 39°C.

Techniques: Flow Cytometry, Marker, Derivative Assay, Cell Culture, Expressing, Positive Control