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John M. Baust

State University of New York - Binghamton, N.Y.

Sponsor: Robert Van Buskirk

"Cell Viability Improves Following Inhibition of Cryopreservation-Induced Apoptosis."

Traditional approaches to cryopreservation of human cells and engineered tissues rely on the addition of molar concentrations of penetrating cryoprotectants contained in either isotonic or hypotonic solutions. This approach rarely considers the ionic balance, buffering capacity or other factors thought to be necessary to avoid hypothermic stress. John Baust’s research in the past few months has shown that HypoThermosol is a superior cryoprotectant to the industry standard. More specifically, cells separately cryopreserved (-196 deg. C ) in either HypoThermosol or 5% DMSO in cell culture media yielded a 30% survival rate; wheras cells cryopreserved in HypoThermosol +5% DMSO had up to a 82% survival rate. These observations suggest that HypoThermosol may be both a superior hypothermic solution (storage at 4 deg C ) as well as a superior cryoprotectant (storage at -196 deg C).

Under IFER funding John Baust plans to conduct the following studies: 1. Test the addition of selected HypoThermosol additives listed in ”Laboratory Research Background” to determine if these additives improve HypoThermosol’s ability to act as cryoprotectant. 2. Test optimized HypoThermosol as a cryoprotectant for the most common cells strains used for product safety testing ( normal human epidermal keratinocytes, human renal cells, human hepatocytes). 3. Isolate the role of apoptosis and necrosis in cryopreservation. 4. Test the optimized HypoThermosol solution for its ability to cryopreserve the engineered human epidermis, EpiDerm (MatTek Corporation, Ashland, MA). The sum consequence of John Baust’s research will be to develop improved techniques for cryopreservation to facilitate the distribution of engineered human cells and tissues for product safety testing.