IADR Abstract Archives
Organoid Models of Normal Human Salivary Glands
Objectives: The objectives of this project were to develop organoid models of human salivary glands, to elucidate the pathogenesis of salivary gland disease and to further understand the development of salivary glands for regenerative studies.
Methods: Organoids were developed from biopsy samples of normal human salivary gland tissue. The cells were isolated and cultured in extracellular matrices (matrigel and/or myogel), as well as on Polymerised High Internal Phase Emulsions (PolyHIPES) scaffolds. They were cultured either at an Air Liquid Interface (ALI) or were submerged in tissue culture medium (non-ALI) and Wnt-3A, R-spondin1, and epidermal growth factor were added to supplement the medium. Haematoxylin and eosin stained sections of the cultures were used to visualise growth. RT-PCR, immunohistochemistry and immunofluorescence were used to determine the differential expression of cell type-specific markers in culture.
Results: Salivary gland cells from single suspensions were able to proliferate and differentiate to form small structures resembling mini-glands for up to 14-days in culture. Organoid structures developed when the cells were grown in matrigel but this was less obvious when grown in myogel. Compared to those grown at an ALI, non-ALI organoids were significantly smaller in size. Cells grown in PolyHIPES scaffolds were able to form budding and branching structures of salivary gland organoids. The organoids showed many characteristics of salivary gland tissue based on their histology and the expression of cell type-specific markers.
Conclusions: Our current data indicates that our culture system enables the growth of human salivary gland organoids. This culture system can be used to study the initial stages of infectious disease and the pathogenesis of other salivary gland diseases.
Methods: Organoids were developed from biopsy samples of normal human salivary gland tissue. The cells were isolated and cultured in extracellular matrices (matrigel and/or myogel), as well as on Polymerised High Internal Phase Emulsions (PolyHIPES) scaffolds. They were cultured either at an Air Liquid Interface (ALI) or were submerged in tissue culture medium (non-ALI) and Wnt-3A, R-spondin1, and epidermal growth factor were added to supplement the medium. Haematoxylin and eosin stained sections of the cultures were used to visualise growth. RT-PCR, immunohistochemistry and immunofluorescence were used to determine the differential expression of cell type-specific markers in culture.
Results: Salivary gland cells from single suspensions were able to proliferate and differentiate to form small structures resembling mini-glands for up to 14-days in culture. Organoid structures developed when the cells were grown in matrigel but this was less obvious when grown in myogel. Compared to those grown at an ALI, non-ALI organoids were significantly smaller in size. Cells grown in PolyHIPES scaffolds were able to form budding and branching structures of salivary gland organoids. The organoids showed many characteristics of salivary gland tissue based on their histology and the expression of cell type-specific markers.
Conclusions: Our current data indicates that our culture system enables the growth of human salivary gland organoids. This culture system can be used to study the initial stages of infectious disease and the pathogenesis of other salivary gland diseases.