Organotypic (raft) culture of biopsy-derived upper respiratory epithelium.
OBJECTIVE
The objective of this study was to derive a reliable technique for culturing biopsy-derived upper respiratory epithelium in a system that supports epithelial differentiation and simulates the normal epithelial life cycle.


METHODS
The authors conducted a prospective study of modification and development of an in vitro tissue culture method.


METHODS
Thirty biopsy specimens from 16 individuals with recurrent respiratory papillomatosis and chronic tonsillitis, pretreated to prevent bacterial and fungal overgrowth, were digested with trypsin to create a supernatant of individual cells. The cells were plated and incubated. At 14 to 16 days, the resulting colonies were placed on a wire cloth raft and fed through diffusion from the underlying culture medium in an air-liquid interface.


RESULTS
Eight specimens were successfully cultured for an average of over 32 days. The longest duration of sustained growth was 60 days. Low-risk human papillomavirus specimen-based cultures reproduced infection in cultured squamous epithelium with corresponding histopathologic features indicating a high level of stratification and differentiation.


CONCLUSIONS
Unlike commercially available cell lines, biopsy-derived material is predisposed to contamination, and successful in vitro culture and experimentation creates many unique challenges. An organotypic culture system, capable of reproducing the differentiation-dependent replication cycle of human papillomavirus, may be used for culturing biopsy-derived specimens for a variety of studies.
Schweinfurth
John Mark
JM
Department of Otolaryngology, University of Mississippi Medical Center, Jackson, Mississippi, USA. JSchweinfurth@ent.unsmed.edu
Meyers
Craig
C
D001706:Biopsy / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media; D004847:Epithelial Cells / Q000166:cytology* / Q000821:virology; D006801:Humans; D027383:Papillomaviridae / Q000254:growth & development* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000821:virology*; D020545:Respiratory Mucosa / Q000166:cytology*; D014069:Tonsillitis / Q000821:virology
D003470:Culture Media