Cultured human nasal epithelial multicellular spheroids: polar cyst-like model tissues

Abstract

We report here a new readily cultured nonadherent hollow spheroidal epithelial tissue model: human nasal epithelial multicellular spheroids, prepared from brushings of human nasal epithelium in vivo. Although cultured cyst-like epithelial models developed from embryonic, transformed, or polypoid tissues have been reported previously, human nasal epithelial multicellular spheroids are derived from normal mature nontransformed human airway epithelial cells. In our studies, spheroids ranged in size from 50 to 700 microns diameter (averaging approximately 250 microns). Cells of the spheroid displayed morphological polarity and formed junctional complexes. Transcellular electrolyte transport may underlie the increase in spheroid size which occurred in culture. The ease and simplicity of the brushing and culture procedures reported here render normal and diseased human cell populations more readily accessible to investigation. We believe human nasal epithelial multicellular spheroids may have important applications in the study of electrolyte and fluid transport processes, ciliary motility, epithelial polarity, cellular metabolism, and drug cytotoxicity in normal and pathophysiological states of the human respiratory tract (e.g., cystic fibrosis).

Polarized Expression of Cystic Fibrosis Transmembrane Conductance Regulator and Associated Epithelial Proteins during the Regeneration of Human Airway Surface Epithelium in Three-Dimensional Culture

“…We did not observe airway cell proliferation even during the first days of 3-D culture. This is in agreement with the data reported by Bridges et al (1991), Jorissen et al (1991), Pedersen et al (1999), and Ulrich et al (1998), which suggest that these cells cease to proliferate as soon as they aggregate and form junctions. During the first 15 days of culture, ciliated cells could not be identified and all spheroid surface epithelial cells were covered with microvilli.…”

“…The diameter of these spheroids averaged 50 to 100 m and remained within this range throughout the course of the study. The morphology of these spheroids is similar to that described by Bridges et al (1991), Jorissen et al (1991), Pedersen et al (1999), and Ulrich et al (1998) and is generally composed of a monolayered or occasionally two-layered epithelium. Percentage of basal CK13-positive cells and CK18-positive cells did not differ significantly before and after 3-D culture.…”

“…Several investigators have previously shown that epithelial sheets, dissociated from adult airway surface epithelium, nasal brushings, or nasal polyps and cultured in suspension spontaneously reform free-floating epithelial spheroids in which ciliary activity and ion transport functions could be maintained for several months (Bridges et al, 1991;Jorissen et al, 1991;Pedersen et al, 1999;Ulrich et al, 1998). In all of these studies, the native well-organized surface epithelial cell structures were preserved and no information could therefore be obtained on the depolarization and repolarization process.…”

Alternative Method for Primary Nasal Epithelial Cell Culture Using Intranasal Brushing and Feasibility for the Study of Epithelial Functions in Allergic Rhinitis

“…Do Yang Park, 1 Sujin Kim, 2 Chang-Hoon Kim, 3,4,5 Joo-Heon Yoon, 3,4,5 Hyun-Jik Kim 4,6 sues are widely used for sampling nasal specimens from subjects. However, these tissues may not replicate the physiologic condition due to the preconditioning process required before the subculture.…”

A murine tracheal culture system to investigate parameters affecting gene therapy for cystic fibrosis

“…23,24 The discrepancy between in vitro and in vivo gene transfer efficiency creates a need for a model system which accurately reflects the difficulties of delivery to the airway epithelium in vivo. Model systems that have been developed include human nasal epithelial multicellular spheroids, 25 human nasal epithelial brushings, the generation of human airway xenografts in rats or immunodeficient mice 26,27 and culture of sheep tracheal explants. 28 We have developed a simple, tracheal culture system and used this to test parameters affecting transfection efficiency which cannot be readily assessed in vivo.…”