Abstract
The biopreservation sciences have typically relied on the utilization of low temperature to suppress or arrest cellular physiological function in an attempt to extend biological time. Recently, however, several reports have detailed the successful anhydrobiotic (dry-state) preservation of cellular systems through the utilization of stabilizing molecules, such as trehalose. In this study, we investigated the influence of cell substrate interaction during and following desiccation on cell survival in a human fibroblast model. Cells were dried on differing culture surfaces (extra-cellular matrices, including poly-L-lysine, collagen, laminin, fibronectin, and standard plasma treated tissue culture ware) after allowing 10 min for loose cell attachment. Desiccated samples (13% residual moisture) were rehydrated and then cultured on the various differing extra-cellular matrices. Samples cultured on biologically active matrices (collagen, fibronectin, and laminin) following desiccation resulted in a significant improvement (>15%) in cell survival in comparison to poly-L-lysine and plasma-treated tissue culture surfaces. Furthermore, both desiccation and the subsequent culture of samples on biologically active surfaces resulted in ,~30% increase in overall cell survival in comparison to standard culture. Our findings indicate that cell culture substrate may play a substantial role in the success of anhydrobiotic preservation. In addition, the data suggests that current methodologies utilized to achieve dry-state preservation may translate to the preservation of more complex biological systems, such as engineered tissue constructs, where the maintenance of functional construct integrity is equally as important as cell survival.
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