Use of Frozen–Thawed Cervical Tissues in the Organ Culture System to Measure Anti-HIV Activities of Candidate Microbicides

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Abstract
Cervical tissue-based organ culture system has been used to test the cytotoxicity and antiviral activity of microbicides. One of the problems of using current organ culture methods for routine microbicide testing is the need to continually obtain fresh tissue, which can be limited in access and supply. Use of frozen tissue, stored when available and thawed when needed, would alleviate the need for constant access to new tissue. This study was designed to explore the possibility of using frozen–thawed cervical tissue to test microbicides for their anti-HIV activity. We provided biochemical, histological, and quantitative immunohistochemical data to demonstrate the integrity of the frozen–thawed organ culture system. Significant levels of HIV-1 mucosal transmission were noted with both fresh and frozen–thawed tissue, regardless of the coreceptor usage of the virus isolate. Furthermore, candidate microbicides UC781, β-cyclodextrin, and octylglycerol inhibited HIV-1 transmission across the mucosa of frozen–thawed tissues with a level of efficiency similar to that of fresh tissues. Therefore, frozen–thawed cervical tissue in the organ culture system provides a practical and convenient model to screen topical microbicides for their ability to block sexual transmission of HIV-1, and reduces the problems associated with procurement of the numerous tissues required for evaluation and comparison of microbicide candidates among different laboratories.

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