Principle of Assay
The organoid culture assay is based on the ability of primary tissue-derived cells to self-organize into three-dimensional structures when provided with an appropriate extracellular matrix and a defined, lineage-specific growth environment. Fresh tissue is first processed using a controlled enzymatic digestion solution that releases viable stem and progenitor cell clusters while preserving surface receptors and cellular polarity. The resulting fragments are embedded in a matrix such as Matrigel, which functions as a scaffold that supports 3D architecture and enables signaling through integrins and basement-membrane interactions. The cultures are then supplied with a specialized organoid medium containing niche-mimicking growth factors, small molecules, and nutrients that maintain stemness while permitting tissue-specific differentiation. During routine expansion, a gentle passaging enzyme is used to fragment mature organoids into smaller units without compromising viability, allowing serial propagation over many passages. For biobanking, a cryopreservation formulation containing permeable and non-permeable cryoprotectants is applied to prevent ice-induced damage and maintain long-term genomic and functional stability. After thawing, a recovery buffer restores isotonic balance and supports rapid re-establishment of 3D growth. Together, this workflow enables the continuous generation, expansion, freezing, and re-culturing of physiologically relevant organoids that retain genetic, phenotypic, and microenvironmental features of the source tissue, making the system suitable for disease modeling, drug screening, and personalized research applications.
