Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

Figure 5

CEDPs differentiation potential. (a) Optical microscope image of a sphere formed in the absence of SB-216763 after 5 days of CEDPs differentiation. (b) cTnT⁺ cells formation after immunostaining with anti-cTnT and anti-VE-cadherin. (c–e) Formation of intercalated disk structures in beating spheres after 10 days of CEDPs differentiation, shown by double-IF-staining with anti-cTnT/anti-Desmoplakin (DSP), anti-cTnT/anti-Desmocollin-2 (DSG2), and anti-MyHC/anti-Desmoplakin. Note the striations of the sarcomeric cTnT staining and the punctate desmosomal staining in areas connecting adjacent cardiomyocytes. (f) Expression of MLC2v and MLC2a after 10 days of CEDPs differentiation by RT-PCR analysis. (g) VE-cadherin⁺ cells in CEDPs-derived differentiated cells. Note extensive adherens junctions formation between endothelial cells. Magnification corresponds to marked area. (h-i) Expression of vWF and CD39 in VE-cadherin⁺ endothelial cells after 10 days of CEDPs differentiation. (j-k) Expression of Isl1 and Mef2c progenitor markers in a-actinin⁺ and VE-cadherin⁺ cells, respectively, at day 10 of CEDPs differentiation. (l) Expression of SMA during CEDPs differentiation. Draq5 counterstained DNA (b, g). Scale bar: 20 μm.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)

(l)