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

Figure 1

Explant organ culture system of neonatal foreskin. (a) Cell migration from skin tissue after 3 days (left); fibroblast-like cells can be observed migrating and sprouting out from tissue after 12 days (right). (b) A transwell migration assay where cells migrated to the lower chamber visualized by Eosin staining. The transwell migration assay was repeated and the total number of cells that migrated was quantified to determine the relative numbers of migrated cells (b, lower panel). Data are presented as mean ± S.D. The experiment was run in triplicate. (c) The real time migration was executed using the xCELLigence RTCA DP device. Cells were seeded per well in 16-well microelectronic sensing, two-chamber transwell plates (CIM-plates). The electrical impedance was captured every 15 min for an experimental duration of ~68 h. The rate of migration is expressed as the CI or the change in electrical impedance at each time-point (c, left). Values are expressed as the ±SEM of the 8 replica wells from three independent experiments (c, right). (d) Measurement of undifferentiated hNSSCs cell migration by in vitro wound healing scratch assay. The migration ability was assessed 12 h and 24 h from injury. The figure shows the migration of undifferentiated cells immediately after the scratch 0 h, 12 h, and 24 h (Bar = 100 μm). (e) ALDH activity of hNSSCs was measured using the Aldefluor assay and FACS analysis. Cells incubated with specific inhibitor of ALDH, DEAB, were used to establish the baseline fluorescence of these cells and to define the Aldefluor positive region.

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