The mission of BioLife Technologies (BLT) is to develop improved hypothermic (Cold storage at 4 - 10 degree C) and cryopreservation (-196 degree C) solutions designed to maintain human cells, tissues and organs in a near state of suspended animation. Data presented herein demonstrate that BLT's solutions, the HypoThermosol (HTS) series, are better at protecting kidney, heart and skin cells than is ViaSpan- a product produced by DuPont. BLT has launched an aggressive program to determine the molecular basis of cell death during extended cryopreservation and hypothermic storage so that knowledge of these events can lead to a new generation of cryopreservation solutions. The Specific Aims of this Phase I project are designed to determine (1) which apoptotic inhibitors are best at protecting the engineered human epidermis, EpiDerm, during cryopreservation and hypothermic storage, (2) if upregulation of the cell death inhibiting protein bcl-2 can protect cells from cryopreservation and hypothermia-induced cell death, and (3) if cytochrome c, an apoptosis trigger, is released from mitochondria in human skin cells that have been preserved. A higher risk Specific Aim is proposed to make cold-tolerant mutants from an E67-tranfected human keratinocyte cell line that can withstand hypothermia and/or cryopreservation regimes better than parental progenitors. Completion of the studies will allow BLT to establish itself as the leader in the preservation of engineered human cells and tissues.
