Novel fixed z-direction (FiZD) kidney primordia and an organoid culture system for time-lapse confocal imaging

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Saarela, Ulla | Akram, Saad Ullah | Desgrange, Audrey | Rak-Raszewska, Aleksandra | Shan, Jingdong | Cereghini, Silvia | Ronkainen, Veli-Pekka | Heikkila, Janne | Skovorodkin, Ilya | Vainio, Seppo J.

Edité par CCSD ; Company of Biologists -

International audience. Tissue, organ and organoid cultures provide suitable models for developmental studies, but our understanding of how the organs are assembled at the single-cell level still remains unclear. We describe here a novel fixed z-direction (FiZD) culture setup that permits high-resolution confocal imaging of organoids and embryonic tissues. In a FiZD culture a permeable membrane compresses the tissues onto a glass coverslip and the spacers adjust the thickness, enabling the tissue to grow for up to 12 days. Thus, the kidney rudiment and the organoids can adjust to the limited z-directional space and yet advance the process of kidney morphogenesis, enabling long-term time-lapse and high-resolution confocal imaging. As the data quality achieved was sufficient for computer-assisted cell segmentation and analysis, the method can be used for studying morphogenesis ex vivo at the level of the single constituent cells of a complex mammalian organogenesis model system.

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