Multipotent stem cell-based culture systems and models

A method of generating a 3-dimensional tissue engineering model comprising the steps of:

• (a) propagating multipotent stem cells from human skin fibroblast culture by growing the cells in a culture containing amniotic fluid growth medium (AFM) and allowing the cells to propagate for at least 3 passages;
• (b) culturing the multipotent stem cells in the laminin-coated bead plates in a tissue culture media that promotes differentiation into one of the three germ layers,
• wherein the laminin-coated bead plates were created by:
• (1) dissolving laminin in cold phosphate buffer saline (PBS) placed in a tissue culture plate;
• (2) adding sterile spherical glass beads or a mix of spherical glass beads to the laminin;
• (3) placing the culture plate in an incubator at 37° C. for at least 12 hours in order to induce polymerization of laminin;
• (4) removing excess PBS and allowing the culture plate to completely air dry;
• (5) adding the multipotent stem cells to the laminin-coated bead plates;
• and
• (6) plating the multipotent stem cells in the laminin-coated bead plates with the multipotent stem cells in an incubator at 37° C.;
• and
• (c) subjecting the multipotent stem cells to lineage-specific differentiation under suitable conditions into cells of any of three germ layers.

A method for identifying one or more genes involved in the process of lineage-specific differentiation, said method comprising the steps of:

• (a) propagating multipotent stem cells from human skin fibroblast culture by growing the cells in a culture containing amniotic fluid growth medium (AFM) and allowing the cells to propagate for at least 3 passages;
• (b) subjecting said multipotent stem cells to lineage-specific differentiation by culturing said multipotent stem cells under culture conditions suitable for lineage-specific differentiation until differentiated cells result;
• (c) subjecting said differentiated cells to gene expression profiling using microarray technology;
• and
• (d) determining which one or more genes is upregulated or downregulated during the process of lineage-specific differentiation.