Dehydration through evaporation, or air drying, is expected to have both similarities and differences to osmostress. Both stresses involve water loss, but the degree of dehydration will ultimately be more severe during desiccation. Despite the severity of desiccation stress, there are examples of organisms that can survive almost complete water loss, including resurrection plants and plant seeds, certain invertebrates among the nematodes, brine shrimps, tardigrades and bdelloid rotifers, and many microorganisms, including bakers' yeast. During desiccation, these organisms enter a state of suspended animation, a process known as anhydrobiosis ("life without water"). For other organisms, desiccation is lethal, but there is considerable interest in using what is known about anhydrobiosis to confer desiccation tolerance on sensitive cell types, such as mammalian cells. Success with this approach, which we have termed anhydrobiotic engineering, will require a more complete knowledge of the mechanisms of desiccation tolerance and the sensing and response of nontolerant organisms to extreme dehydration. With this goal in mind, we have attempted to characterize the response of human tissue culture cells to desiccation and to compare this response with osmotic upshift. This chapter describes some of the methods used to begin to uncover the response to evaporative water loss in human cell cultures.