With the advancements being made in tissue engineering, cell transplantation, stem cell biology, and gene therapy, the clinical demand for effective long-term storage methods for cells and tissues will continue to increase. We propose to develop novel methods to biostabilization of mammalian cells for long-term preservation in a desiccated state at ambient temperature. In nature, many animals and organisms down regulate their metabolism and may enter into a state of stasis by either desiccation through removal of water from their cells (i.e., anhydrobiosis) or by a developmentally programmed arrest under full hydration (i.e., diapause). The ability to enter diapause prior to desiccation is crucial for the survivorship of many organisms that undergo natural states of dormancy. Furthermore, a common theme is that desiccation-tolerant animals accumulate large amounts of disaccharides, especially trehalose and sucrose. These sugars provide protective effects by forming stable sugar glasses at high water contents, and by stabilizing biological membranes and proteins through direct interaction with polar residues. We, therefore, hypothesize that metabolic preconditioning of mammalian cells to induce diapause-like state followed by controlled drying, storage, and rehydration conditions (i.e., physicochemical, biochemical, and metabolic) can be used to achieve desiccation tolerance in mammalian cells and tissues. To this end, our 3 distinct, but interactive, specific aims are: 1) To develop optimal physicochemical conditions to stabilize desiccated cells. 2) To metabolically precondition mammalian cells to improve survivorship during storage. 3) To develop metabolic and biophysical strategies to accelerate recovery of desiccated cells. This project is one of the first attempts to apply engineering and quantitative concepts to achieving anhydrobiotic state in mammalian cells. It provides a systems view of the metabolic and cellular changes a cell encounters before, during, and after desiccation. This project is inspired by nature and it uses engineering concepts and approaches to translate nature's solution to long-term storage or """"""""suspended animation"""""""" for mammalian systems. The proposed studies will significantly impact on human health by providing a solution to the problem of providing long-term storage of blood cells, stem cells, tissue engineered products, and cell-based biosensors for use in regenerative medicine, tissue engineering, and bioterrorism. In the short term, it will help increase the treatment modalities available to liver failure by providing stable, long-term stabilized cells for bioartificial liver assist devices. The longer-range outcome of the proposed research is to translate the information gained from these studies into whole organ preservation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM071345-01
Application #
6796449
Study Section
Special Emphasis Panel (ZRG1-SSS-U (10))
Program Officer
Anderson, James J
Project Start
2004-08-01
Project End
2008-07-31
Budget Start
2004-08-01
Budget End
2005-07-31
Support Year
1
Fiscal Year
2004
Total Cost
$480,833
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Hand, Steven C; Menze, Michael A; Borcar, Apu et al. (2011) Metabolic restructuring during energy-limited states: insights from Artemia franciscana embryos and other animals. J Insect Physiol 57:584-94
Menze, Michael A; Fortner, Grady; Nag, Suman et al. (2010) Mechanisms of apoptosis in Crustacea: What conditions induce versus suppress cell death? Apoptosis 15:293-312
Reynolds, Julie A; Hand, Steven C (2009) Decoupling development and energy flow during embryonic diapause in the cricket, Allonemobius socius. J Exp Biol 212:2065-74
Roach, Kenneth L; King, Kevin R; Uygun, Korkut et al. (2009) High-throughput single cell arrays as a novel tool in biopreservation. Cryobiology 58:315-21
Reynolds, Julie A; Hand, Steven C (2009) Embryonic diapause highlighted by differential expression of mRNAs for ecdysteroidogenesis, transcription and lipid sparing in the cricket Allonemobius socius. J Exp Biol 212:2075-84
Holman, Jeremy D; Hand, Steven C (2009) Metabolic Depression is Delayed and Mitochondrial Impairment Averted during Prolonged Anoxia in the ghost shrimp, Lepidophthalmus louisianensis (Schmitt, 1935). J Exp Mar Bio Ecol 376:85-93
He, Xiaoming; Fowler, Alex; Menze, Michael et al. (2008) Desiccation kinetics and biothermodynamics of glass forming trehalose solutions in thin films. Ann Biomed Eng 36:1428-39
Hand, Steven C; Menze, Michael A (2008) Mitochondria in energy-limited states: mechanisms that blunt the signaling of cell death. J Exp Biol 211:1829-40
Hand, Steven C; Jones, Dana; Menze, Michael A et al. (2007) Life without water: expression of plant LEA genes by an anhydrobiotic arthropod. J Exp Zool A Ecol Genet Physiol 307:62-6
Covi, Joseph A; Hand, Steven C (2007) Energizing an invertebrate embryo: bafilomycin-dependent respiration and the metabolic cost of proton pumping by the V-ATPase. Physiol Biochem Zool 80:422-32

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