Example 1
The pluripotent stem cell line H9 was obtained from NIH line WA 09, supplied by WiCell (Madison, Wis.) and was maintained in an undifferentiated state by passaging on irradiated human foreskin fibroblasts (line HS27, ATCC, Manassas, Va.) and gelatin coated plates. To differentiate the pluripotent stem cells towards a mesodermal and then mesenchymal lineage, the colonies of the pluripotent stem cells were mechanically dissected into small pieces under microscopic guidance and then transferred to tissue culture-treated 6-well plates (Corning). The cells at this stage were considered passage 0 (P0). The cells were cultured in DMEM/F12 supplemented with non-essential amino acids and 10% fetal bovine serum (FBS, Invitrogen-Gibco, Grand Island, N.Y.). When the culture approached confluency, cells were trypsinized and transferred to a new tissue culture flask.
