In vitro 3D cultures have opened up new horizons for translational human disease research, disease modeling, regenerative medicine and predictive precision therapies. Today, differentiated organ cell types can be generated in vitro from variety of stem cells (SCs) to mimic complex tissues while retaining genomic stability and response to drug treatments.
Thus, organoids have a distinct advantage over traditional 2D cultures and hold unprecedented potential for various, innovative applications. However, in order to exploit these models for meaningful basic research, disease modeling and drug screening, specific and reliable culture and analysis methods are required. Currently, characterization and optimization of organoid cultures are limited in their ability to reproducibly form and monitor these 3D structures as they form and grow overtime.
The Incucyte® Organoid Culture QC Application is a turnkey solution consisting of a validated protocol and software module for real-time visualization and label-free quantification to optimize organoid culture conditions. Make informed decisions about organoid culture status and open a new world of analysis of advanced cell models.
Generate standardized and reproducible organoid data using live-cell imaging and analysis over days, weeks or months over multiple tissue samples.
Accelerate your organoid discovery with an established protocol and integrated analysis, enabling reproducible culture techniques for downstream assay preparation. The Organoid Culture QC Application allows for direct, label-free analysis of organoid size, count and morphology in Matrigel® domes in a physiologically relevant environment using the Incucyte® Organoid Analysis Software Module and Incucyte® Live-Cell Analysis System.
Locate and Analyze Organoids Automatically
Visualize and quantify organoids embedded within Matrigel® domes automatically in a physiologically relevant environment.
Characterize Organoid Formation Kinetically
Obtain continuous, label-free data to optimize culture conditions using the Incucyte® Organoid Analysis Software Module.
Identify Maturation Phase of Organoid cultures
Define an optimal period for passaging organoids or extension of cultures, based on integrated morphology metrics.
Enable scientific discovery using an established protocol and integrated analysis of organoids in real time.
Figure 1. Kinetically monitor and quantify organoid differentiation and growth in Matrigel® domes undisturbed inside your incubator. Mouse intestinal and mouse pancreatic organoids were embedded in Matrigel® domes (50 or 100% Matrigel® respectively) in 24-well plates and imaged over 6 - 8 days in an Incucyte. Brightfield videos (2 d post seeding) and time-course profiles of brightfield area show cell type specific organoid differentiation, morphology and growth respectively.
Automatically analyse organoid size and count of the entire Matrigel® dome over weeks or months to optimize culture conditions using the Incucyte® Organoid Analysis Software Module.
Figure 2. Distinguish suitable conditions for maximal culture expansion using brightfield size measurements. Mouse hepatic organoids were embedded in Matrigel domes (100%) in 48-well plates at multiple seeding densities. Images (5 d post seeding) and time-courses demonstrate that organoid size and growth rate is directly proportional to cell number.
Use unbiased assessment of organoid morphology to provide insight into optimal period for passaging or extending cultures.
Figure 3. Define cell-type specific passaging frequency using integrated morphology metrics. Intestinal organoid passaging frequency is driven by indicators of maturation such as budding and accumulation of debris within the lumen. Kinetically quantify these indicators using Incucyte’s Eccentricity (budding) or Darkness (debris accumulation) metrics.
Leverage an established protocol and integrated software for reproducible formation and analysis of organoids embedded in Matrigel® domes in a 24- or 48-well plates.
Figure 4. Incucyte lab tested protocol for culturing Organoids in Matrigel Domes. Easily culture, expand and maintain organoid cultures embedded in Matrigel domes for representation of organ specific physiology.
Figure 5. Guided interface is easy to use for even first-time users. Automated image acquisition and analysis tools provide a ‘set up and walk away’ experience. Visualize organoid differentiation and growth in 24-well or 48-well plates remotely to monitor experimental progress and analyse in real time.