A self-contained anaerobic environment-generating culture device is provided. The culture device includes a first substrate having opposing inner and outer surfaces, a second substrate having opposing inner and outer surfaces, a growth region disposed between the inner surfaces of the first and second substrates, an effective amount of a substantially dry enzyme component of an enzyme-mediated oxygen depletion system, an effective amount of a substantially dry enzyme substrate component of the enzyme-mediated oxygen depletion system, and a dry, cold-water-soluble gelling agent disposed in the growth region. The enzyme and enzyme substrate components are disposed in coatings in the growth region. The first and second substrates are substantially nontransmissible to gaseous oxygen. Methods of making and using the culture device area also provided.