Engineering a Human Organ for Dry Storage

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DESCRIPTION (provided by applicant): Tissue engineering is a field with tremendous untapped potential, current revenues ($232 million in 2000, Genetic Engineering News 9/15/01) fall far short of the estimated market of $100 billion (Barron's, 5/15/00). This gap is in large part due to the significant barriers of short shelf life and high storage cost. Stratatech proposes to develop technology overcoming these hurdles using genetic manipulations of source cells coupled with recent advances in preservation solution engineering to create methods enabling room temperature storage of living cells and organs. Currently tissues are preserved by slow freezing during which they suffer numerous damaging effects. Rapid freezing or vitrification avoids these issues by preserving samples in an amorphous glass, but vitrification can give rise to lethal intracellular ice formation. Agents such as trehalose, a disaccharide of glucose, enhance the vitrification properties of preservation solutions; unfortunately, trehalose cannot penetrate cell membranes so novel methods must be used to deliver intracellular trehalose. Furthermore, until recently, vitrification of medically useful sample sized was not practical, but recent work has shown that careful engineering of the preservation solution can greatly enhance trehalose's ability to raise glass transition temperatures. Stratatech is developing a line of human keratinocytes for therapeutic applications called NIKS cells. NIKS cells are an ideal vehicle for this proposal in that they can be perpetually cultured in vitro and they can be used to manufacture a functional human organ. In addition they are pathogen-free, non-tumorigenic and they recapitulate the correct architecture of human skin. NIKS cells and skin tissue grown from them are capable of stable genetic modifications making it possible to optimize their survival in the dry state.