Embryonic stem cells (ESC) have been explored as tools for
studying development, as well as, potential sources for a large number
of therapies in regenerative medicine. Traditionally, ESC are cultured
on tissue culture plastic, however; it has been recently shown that the
stiffness of the environmental substrate can direct the cells towards
various cell lineages. Our laboratory is specifically interested in examining
the combined roles of biochemical and physical signaling in
cardiac and vascular cell fate and patterning these vascular cells into
vascular branch-like tress.Using our novel mouse ESC that expresses a
GFP reporter under the Tie-2 and an RFP reporter under alpha smooth
muscle actin and serum-free inductionmediums, we examined the role
of stiffness in the diverging fate of Flk-1 + vascular progenitor cells.
The results indicate that both of the Flk-1 + vascular progenitor cells
and human umbilical vein endothelial cells (HUVEC) preferentially
adhere to 10 kPa compared with the 1 kPa, and 34 kPa compared with
the 10 kPa. We also observed both the GFP/Tie-2 + endothelial-like
and RFP + smooth muscle-like cells outgrowths from the Flk-1 +
cells, with the GFP/Tie-2 + cells dominating the cultures, supporting
the role of stiffness in vascular fate. Next, we generated a vascular
fractal-like pattern reverse mold and have stamped fibronectin vascular
pattern onto non-tissue culture treated plastic. Using these combined
technologies, we have been able to genera