Angiogenic Potential of Human Neonatal Foreskin Stromal Cells in the Chick Embryo Chorioallantoic Membrane Model

Figure 3

hNSSCs contribute to neovasculature using ex vivo CAM model. (a) Schematic illustration of the organotypic culture system (liquid-air interface). hNSSCs pellets were placed on the top of a confetti that is in contact with the membrane on the insert. The media diffuse across the insert membrane and confetti to reach the pellet. Microscopic aspect of chick chorioallantoic membrane (CAM) and organotypic cultured hNSSCs with confetti grafted to it. (b) hNSSCs pellet placed on top of a piece of confetti that is in contact with the membrane on the insert. (c) Window (2 cm²) made in the egg shell of a 10-day-old chick embryo with organotypic culture transplanted. (d) After 10 days, implantation shell was opened and blood vessel points were observed; (e and f) note the wheel-spoke pattern of CAM blood vessels around the graft (undifferentiated and differentiated). Stimulation of angiogenesis in CAM by undifferentiated or differentiated hNSSCs. The expansion of new blood vessels was determined by counting branch points after 10 days of cell transplantation. Sprouting and branching vessels are prominent in the CAM transplanted by differentiated hNSSCs compared to the undifferentiated hNSSCs transplantation. (g) Quantification of new blood vessels formed by naive (undifferentiated) and differentiated hNSSCs compared to negative control (confetti without cell transplantation) (). . (h, i) Paraffin sections of both undifferentiated and differentiated hNSSCs transplanted CAM were stained by haematoxylin and eosin stain to show the vascular density in CAM; blood vessels were counted again (blue colour points indicate vasculature) with Aperio’s ImageScope software (Aperio Technologies, Vista, CA, USA) (Bar = 100 μm).