Monolayer and three-dimensional cell culture and living tissue culture of gallbladder epithelium

Abstract

Several models for preparing and isolating human and animal gallbladder epithelial cells, including low‐grade gallbladder carcinoma cells, as well as proposed systems for culturing these isolated epithelial cells are reviewed here. Several reports concerning tissue culture of the gallbladder are also reviewed. The cell culture systems are divided into monolayer cell culture on collagen‐coated or uncoated culture dishes or other culture substrate and three‐dimensional cell culture in collagen gel. To prepare and isolate gallbladder epithelial cells, digestion of the gallbladder mucosa, abrasion of the mucosal epithelial cells, and excision of epithelial outgrowth of mucosal explants are applied. In monolayer cell culture, most of the specific biological features of isolated and cultured cells characteristic to the gallbladder are gradually lost after several passages, though quantitative and objective analyses of the pathophysiology of cultured cells and their secretory substances can be performed. Tissue culture using explants of the gallbladder has mainly been used for physiological studies of the gallbladder, such as investigating the transport of water and electrolytes. In this tissue culture system, quantitative assessment is difficult, though the original and specific biological and histological characteristics of the gallbladder are retained. Three‐dimensional collagen gel culture could be an ideal model combining monolayer cell culture and tissue culture systems, and create controllable conditions or environments when several biologically active substances, such as growth factors, proinflammatory cytokines and adhesion molecules, are added to the culture medium. Advantages and shortcomings of individual cultivation models are discussed, and selecting the culture model most appropriate to the purpose of the study will facilitate investigations of the biology and pathogenetic mechanisms of gallbladder diseases such as cholelithiasis. Microsc. Res. Tech. 39:71–84, 1997. © 1997 Wiley‐Liss, Inc.

Interstitial Cajal-like cells in human gallbladder

“…However, it is worthwhile to mention that human and murine ICLC were similarly distributed, mainly in the sub epithelial and muscular layers. An overview of literature shows that, when attempts were made to culture in vitro gallbladder tissues, little attention was paid to interstitial cells (for details see Nakanuma et al 1997).…”

The role of E-cadherin in the differentiation of gallbladder cancer cells

“…Normal rabbit gallbladder epithelial cells primarily cultured in the collagen gel matrix form monolayered spherical cystic structures. These cells can completely reestablish polarity, showing microvilli on the apical surface, basal lamina on the basal site, and a completed junctional complex at the upper lateral site, and they tightly adhere to neighboring cells (Yoshida et al 1996;Nakanuma et al 1997;. Since the spherical cystic structures formed by GB-d1 cells in SC were microscopically and electron-microscopically similar to those formed by normal rabbit gallbladder cells, it can be speculated that GB-d1 cells in SC behave like moderately differentiated or well-differentiated cancer cells.…”

Lipopolysaccharide Induces Overexpression of MUC2 and MUC5AC in Cultured Biliary Epithelial Cells

“…Murine intrahepatic BECs were isolated from 8-weekold female BALB/c mice and were purified and cultured, as described previously. 21,22 The purified BECs were seeded at a density of 1 ϫ 10 5 /cm 2 on a collagen-coated dish, and were incubated with a culture medium composed of D-MDM/F-12 (Dulbecco's modified Eagle medium and nutrient mixture F-12, 1:1; Life Technologies, Inc., Rockville, MD), 10% Nu-Serum (Becton Dickinson Labware, Bedford, MA), 1% ITSϩ (Becton Dickinson Labware), 5 mol/L forskolin (Wako Pure Chemical Industries, Ltd., Osaka, Japan), 12.5 mg/ml of bovine pituitary extract (Life Technologies, Inc.), 1 mol/L dexamethasone (Sigma Chemical Co., St Louis, MO), 5 mol/L triiodothyronine (Sigma Chemical Co.), 5 mg/ml of glucose (Sigma Chemical Co.), 25 mmol/L sodium bicarbonate (Sigma Chemical Co.), 1% antibiotics-antimycotic (Life Technologies, Inc.), and 25 ng/ml of mouse epidermal growth factor (Life Technologies, Inc.) at 37°C in an atmosphere of 5% CO 2 for 2 weeks. The cells formed a monolayer with a sheet-like growth pattern.…”