Self-Organized Vascular Networks from Human Pluripotent Stem Cells in a Synthetic Matrix

A method for differentiating mammalian pluripotent stem cells (PSCs) towards a population that can self-organize into vascular networks in vitro, comprising the steps of:

• plating a single-cell suspension of PSCs onto a suitable surface;
• adding culture medium
• culturing the cells for several days;
• adding VEGF and a transforming growth factor-beta (TGF-β) inhibitor to the culture medium;
• culturing the cells for several days;
• harvesting the resulting cells;
• and
• embedding the harvested cells in a hydrophilic matrix.

A method for differentiating mammalian pluripotent stem cells (PSCs) into early epithelial cells (ECs) in vitro, comprising the steps of:

• plating a single-cell suspension of PSCs onto a suitable surface;
• adding culture medium
• culturing the cells for several days;
• adding VEGF and a transforming growth factor-beta (TGF-β) inhibitor to the culture medium;
• culturing the cells for several days;
• and
• sub-culturing the resulting EVCs or sub-culturing VEcad+ cells sorted from the EVCs in a medium containing VEGF and TGF-β inhibitor.

A method for differentiating mammalian pluripotent stem cells (PSCs) into pericyte cells in vitro, comprising the steps of:

• plating a single-cell suspension of PSCs onto a suitable surface;
• adding culture medium
• culturing the cells for several days;
• adding VEGF and a transforming growth factor-beta (TGF-beta inhibitors-β) inhibitor to the culture medium;
• culturing the cells for several days;
• harvesting the resulting EVCs;
• and
• sub-culturing EVCs in a medium containing serum on a tissue-culture treated surface and removing unattached cells after 4 hours.