Tracheal remodeling: comparison of different composite cultures consisting of human respiratory epithelial cells and human chondrocytes

Abstract

The reconstruction of extensive tracheal defects is still an unsolved challenge for thoracic surgery. Tissue engineering is a promising possibility to solve this problem through the generation of an autologous tracheal replacement from patients' own tissue. Therefore, this study investigated the potential of three different coculture systems, combining human respiratory epithelial cells and human chondrocytes. The coculture systems were analyzed by histological staining with alcian blue, immunohistochemical staining with the antibodies, 34betaE12 and CD44v6, and scanning electron microscopy. The first composite culture consisted of human respiratory epithelial cells seeded on human high-density chondrocyte pellets. For the second system, we used native articular cartilage chips as base for the respiratory epithelial cells. The third system consisted of a collagen membrane, seeded with respiratory epithelial cells and human chondrocytes onto different sides of the membrane, which achieved the most promising results. In combination with an air-liquid interface system and fibroblast-conditioned medium, an extended epithelial multilayer with differentiated epithelial cells could be generated. Our results suggest that at least three factors are necessary for the development towards a tracheal replacement: (1) a basal lamina equivalent, consisting of collagen fibers for cell-cell interaction and cell polarization, (2) extracellular factors of mesenchymal fibroblasts, and (3) the presence of an air-liquid interface system for proliferation and differentiation of the epithelial cells.

Tissue engineering and the use of stem/progenitor cells for airway epithelium repair

“…Whether fibroblasts themselves or products produced by fibroblasts are required is not yet resolved. Airway epithelial cells should be co-cultured in a system that includes a basal lamina equivalent, extracellular factors of mesenchymal fibroblasts, and the presence of an air-liquid interface system for proliferation and differentiation of the epithelial cells (MacPherson et al, 2005;Pfenninger et al, 2007). The presence of fibroblasts markedly speeds up the growth of epithelial cells Kobayashi et al, 2007).…”

Overview of Tracheal Tissue Engineering: Clinical Need Drives the Laboratory Approach

“…One study evaluated the performance of (1) epithelial cells on chondrocyte pellets, (2) epithelial cells on native cartilage explants, and (3) co-culture on separate sides of collagen membranes. 66 Immunohistochemical staining revealed expansion and differentiation in the epithelial layer of the third method. The authors of this study concluded that the culture conditions of the third method (collagen membrane system)-a basal lamina equivalent, fibroblast conditioned medium, and airliquid interface-were required for epithelial cell proliferation and differentiation.…”

“…The authors of this study concluded that the culture conditions of the third method (collagen membrane system)-a basal lamina equivalent, fibroblast conditioned medium, and airliquid interface-were required for epithelial cell proliferation and differentiation. 66 Another group cultured human respiratory epithelial cells and chondrocytes separately on 3D scaffolds, and then fused the two constructs together with fibrin surgical adhesive. After 5 days in culture, the constructs remained viable and the layers were appropriately adhered to each other.…”

Tracheal Tissue Engineering

“…Prior to adding the cells to the scaffolding, researchers have cultured the cells together or separately. One study evaluated the performance of epithelial cells on chondrocyte pellets, epithelial cells on native cartilage explants, and coculture on opposite sides of collagen membranes [168]. Immunostaining staining revealed expansion and differentiation in the epithelial layer only is the final method.…”