Cryopreservation and glycerol preservation are 2 successful methods for long-term preservation of human cadaver skin. Preservation is subjected to strict criteria to minimize the risk of disease transmission. This investigation compares the effects of glycerol preservation and cryopreservation on the inactivation of HIV-1. The effects of glycerol preservation and cryopreservation on inactivation of both extracellular and intracellular HIV-1Ba-L were investigated. After exposing HIV-1Ba-L-infected material to various concentrations of glycerol or to 10% dimethyl sulfoxide followed by cryopreservation, uninfected peripheral blood mononuclear cells were added to the treated material. At different time points during the culture, supernatants were taken to quantify HIV-1Ba-L and reverse transcriptase levels to determine HIV-1Ba-L infectivity. Cell-free HIV-1Ba-L was inactivated within 30 minutes in 70% and 85% glycerol. Also, intracellular HIV-1Ba-L in infected peripheral blood mononuclear cells or infected cadaver skin was completely inactivated by glycerol treatment in vitro. Cryopreservation did not show any extracellular or intracellular HIV-1Ba-L inactivation. Glycerol preservation–but not cryopreservation–of human cadaveric donor skin can inactivate both extracellular and intracellular HIV-1.