Mature specimens have been used in the traditional or folk medicines of China, Japan, and the Ojibwe as a hemostatic or wound dressing agent.

Other species they included in this stirps were C. umbrina, C. diguetti, C. lycoperdoides, C. rubroflava, C. ochrogleba, C. excipuliformis (since transferred by some authorities to Handkea), and C. elata.

[10] The specific epithet craniiformis derives from the Ancient Greek words cranion, meaning "brain", and forma, "a form".

[5] The rhizomorphs are well developed, and when cut into longitudinal section, reveal three distinct tissues: an outer cortex, a subcortical layer, and a central core.

[13] The thin and fragile exoperidium (the outer layer of "skin") is whitish-gray to gray, and initially smooth before becoming areolate (divided by cracks into discrete areas).

[12] C. bicolor is a smaller, rounder puffball that could be confused with younger specimens of C. craniiformis, but the former species has more coarsely ornamented spores, and lacks a distinct subgleba.

[16] Handkea utriformis is roughly similar in appearance to C. craniiformis, but unlike the latter it develops a cavernous opening to reveal an olive-brown gleba, and has distinct slits in its capillitial threads.

[12] Early 20th-century mycologist Charles McIlvaine noted over a century ago that "the slightest change to yellow makes it bitter.

[20] In the United States, the Ojibwe used the powdery gleba as a hemostatic agent to staunch the flow of nosebleeds:[21] the spore powder was inhaled through the nostrils.

[23] Although Calvatia craniiformis is generally considered a saprobic species, in controlled laboratory conditions, an ectomycorrhizae between the fungus and American sweetgum (Liquidambar styraciflua) was reported in a 1966 publication.

A Chinese study showed that C. craniifromis would readily form mycorrhiza with poplar seedlings on unsterilized, but not on sterilized soil.

[25] Brain puffballs grow singly or in groups in fields and open woods, hardwood forests, and wet areas.

[33] Extracts of the puffball have strong antitumor activity in mouse models attributable to protein-bound polysaccharides, the compounds calvatan, craniformin, and a tautomer of rubroflavin.