Journal: Journal of Virology

Article Title: Development of a Primary Human Cell Model for the Study of Human Cytomegalovirus Replication and Spread within Salivary Epithelium

doi: 10.1128/JVI.01608-18

Figure Lengend Snippet: Establishment of salispheres and primary epithelial lines from human salivary gland. (A) Freshly isolated salivary gland tissues (top) were digested with collagenase-dispase, and single-cell suspensions were then cultured on Cultrex PathClear basement membrane extract (BME) gels, where multicellular salispheres developed within 72 to 96 h (middle). Salispheres were then dissociated, counted, and replated onto Cultrex PathClear BME gels as in the middle panel or plated onto plastic culture dishes to facilitate HCMV infection experiments (bottom). (B) Salispheres were used for mRNA preparation and amylase 1 and hypoxanthine-guanine phosphotransferase (HPRT) expression. (C) Salispheres were then used for more-detailed analyses of salivary gland-specific gene expression. Taken together, the results indicate that the salispheres grown in vitro exhibit gene expression profiles similar to those of salivary tissue in vivo.

Article Snippet: Following digestion and lysis of red blood cells, the salivary cells were plated on Cultrex PathClear basement membrane extract (BME) gels (R&D Systems) and cultured in BEGM growth medium (containing the BEGM bullet kit and 4% charcoal-stripped fetal bovine serum [FBS]), provided by Lonza.

Techniques: Isolation, Cell Culture, Membrane, Infection, Expressing, Gene Expression, In Vitro, In Vivo