Histoculture and Infection with HIV of Functional Human Lymphoid Tissue on Gelfoam®

Abstract

Gelfoam^® histoculture provides a valuable tool for experimental studies of normal and pathological tissue physiology. It allows us to understand cell-cell interactions by mirroring their original spatial relationship within body tissues. Gelfoam^® histoculture can be employed to model host-pathogen interactions mimicking in vivo conditions in vitro. In the present chapter, we describe a protocol to process and infect lymphoid tissue explants with HIV and maintain them in Gelfoam^® histoculture at the liquid-air interface. The Gelfoam^® histocultures with human immunodeficiency virus (HIV) type 1-infected tissues have been used to further understand the biology of early HIV-1 pathogenesis, as well as a novel ex vivo platform to test the efficacy and toxicity of antiviral drugs.

Keywords: Antibody production; Gelfoam®; HIV-1 infection; Histoculture; Lymphoid tissue; Tonsils.

Fig 1:. Human tonsil histoculture preparation.

Tonsils are obtained from tonsillectomy or tonsillotomy. Indicated are the areas that should be removed before proceeding with tissue dissection: residual fibroid tissue from the capsule surrounding the tonsil (white), cauterized tissue (brown-black), and bloody/inflamed tissue (red). Also, parts with brown-greenish color that is indicative of tissue necrosis, should be removed.

Fig 2:. Dissection and culture of human tonsillar tissue.

Pieces of tonsils cleared from unwanted parts (e.g. fibroid, cauterized, bloody, necrotic as indicated in Figure 1) were dissected into strips (upper left) and next into blocks of approximately 8 cubic mm (upper right). Tonsillar tissue explants on top of collagen sponges (9 explants/sponge) in a 6-well plate containing culture medium (bottom).