WILLIAM D VEAZEY
Nursing at 62 Loop, Ocala, FL

2003017, Sep 11, 2003

Nov 13, 2002

10/292797

William Veazey - Gainesville FL, US
Karen Moore - Gainesville FL, US

A61K045/00, A61F002/00

424/423000, 424/093700

The delivery of tissue engineering media using an airbrush. William S. Veazey, D M D Nov. 7, 2002 Procedure: Bovine dermal fibroblasts were delivered into a 6-well tissue culture plate using a Badger 100 G airbrush having fine, medium and large nozzles at air pressures 6, 8, 10, 14 and 18 psi. Cells were delivered in a concentration of 200,000 cells per 1 mL of media; 1 mL of this cell suspension was delivered for each repetition. The cells were washed in Hanks Balanced Salt Solution (HBSS) or Phosphate Buffered Saline (PBS). Trypsin (0.25%) was used to detach the cells from the cell culture plate before delivery. Cells were re-suspended in a 50%-50% Dulbecco's Modified Eagle's Medium (DMEM) and F-12 media (one actually purchases the media in this concentration, called DMEM/F12) with 10% Fetal Bovine Serum (FBS), 2 mM L-Glutamine and 0.1 mM 2-Me. It should be stated that cells could be delivered in a variety of media, and one skilled in the art recognizes that one could use HBSS or PBS alone, or could use DMEM/F12 alone, or any other type of vital media to deliver cells. Cell vitality was measured using a trypan blue exclusion assay. Thickness of deposition was not measured. The 1 mL of airbrush delivered media spread in the tissue culture well as media would normally do.