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Advanced Tools for Reduced Variability in Disease Modeling

Advanced Cell Models for Scalable and Reproducible Disease Models

Advanced cell-based approaches, such as 3D cell culture and 2D stem cell models, are increasingly used to create models of human diseases, allowing researchers to study disease development, progression and underlying pathogenesis in a controlled environment.

3D cell culture models, like disease-specific organoids, offer great advantages over traditional 2D culture and/ or animal models as they better recapitulate organ tissue structure and behavior. 

Due to their potential to better mimic patient-specific phenotypes, induced pluripotent stem cells (iPSCs) are excellent sources from which to generate humanized disease models in large quantities. They offer an avenue for building iPSC-derived organoid models of tissue diseases, genetic disorders (e.g., retinitis pigmentosa), infectious diseases (e.g., respiratory, gastrointestinal), degenerative diseases (e.g., cystic kidney disease, Alzheimer’s disease) and cancer. 
 

Related Webinar: Maximizing the Success of 3D Cell Models for Clinical Research

The Promises and Pitfalls of Advanced Cell-based Disease Modeling

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Advantages

  • Reduced animal testing 
  • Better biological relevance – closing translational gap 
  • No interspecies variations
  • Time and cost saving
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Challenges

  • Complexity of human diseases: Capturing genetics and epigenetics can be difficult to achieve.
  • Complexity of workflows: 3D cell models require sophisticated cell culture and characterization methods.
  • Reproducibility: Numerous models and lack of standardization can lead to experimental differences in results. 
  • Scaling up 3D cultures and models for screening and further downstream applications.

Advanced Tools for Reduced Variability in Disease Modeling

RUO Cytokines Cell Culture vials

RUO Growth Factors and Cytokines

Cytokines and growth factors are key immune system signaling molecules that regulate immune responses, inflammation, cell growth, and differentiation. Their research has advanced our understanding of immune-related diseases and led to the development of targeted therapies and vaccines.

Sartorius produces high-quality, recombinant cytokines and growth factors for research purposes, free from animal-derived components. These rigorously tested products ensure reliable results, consistent supply, and meet the highest purity and efficacy standards, with ISO9001-certified quality management.

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Classical Media, Reagents and Supplements

Sartorius media, reagents and supplements are designed to support the growth and maintenance of a variety of cells and cell lines while meeting the highest quality standards. Each lot is manufactured under a strictly controlled process according to a Product Master Record for lot-to-lot consistency. 

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Incucyte® Live-Cell Analysis System

The Incucyte® Live-Cell Analysis System enables real-time live-cell analysis directly inside your incubator. 

Monitor and characterize phenotypic or functional changes of 3D and 2D cell cultures in a physiologically relevant environment, in real time, without the need to perform labeling.  Incucyte® software makes the process of acquiring, viewing, analyzing and sharing images of living cells easier than ever before. 

  • 3D cell model analysis: Organoid assays
  • Tumor spheroid assays
  • Organoid culture QC

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Borghi, R.; Magliocca, V.; Petrini, S.; Conti, L.A.; Moreno, S.; Bertini, E.; Tartaglia, M.; Compagnucci, C. Dissecting the Role of PCDH19 in Clustering Epilepsy by Exploiting Patient-Specific Models of Neurogenesis. J. Clin. Med. 2021, 10, 2754. https://doi.org/10.3390/jcm10132754
 

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Cell Selection Retrieval instrument cellcelector flex

CellCelector Automated Cell Selection and Retrieval Platform

The CellCelector Flex Platform is a fully automated cell imaging and picking system developed for screening, selection and isolation of single cells, clusters, spheroids and organoids as well as single-cell clones and adherent colonies. 

  • Automated scanning, detection and gating of complex 3D structures
  • Organoid transfer with exceptionally low (1 μL) injection volumes of surrounding media into either 100% hydrogel, liquid or any other medium
  • Successful embedding of spheroids and organoids in 100% Matrigel® into plates with or without cell culture membranes

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iQue® Advanced Flow Cytometry Platform

The iQue® Advanced Flow Cytometry Platform is a high-throughput suspension cell and bead analysis platform for rapidly profiling of immune-cell phenotype and function in drug discovery and development workflows. Ideal for screens where cells are precious or limited in number, iQue® is the fastest way to generate high-content data from small samples. 

Microfluidics acquisition capability analyses samples as small as 10 μL in a 384-well format, with zero dead volume. Cell detection occurs at rates of thousands of cells per second.

  • Evaluation of T cell response in advanced 3D tumor models
  • Organoid characterization
  • Immune cell phenotype and function in advanced cell models

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Figure 2 - Schematic Outlining the Protocols for Analysis of ICK in 3D Tumor Spheroids Using the iQue® Platform

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Microsart® Mycoplasma, Bacteria and Fungi Rapid qPCR Test Kits

Testing cell cultures for microbial contamination on a regular basis ensures culture performance and parameters remain consistent and reproducible.

Microsart® Mycoplasma, Bacteria and Fungi qPCR kits offer rapid, reliable and easy-to-use solutions for microbial contamination control in compliance with international guidelines. Get results in just three hours! 

The combination of speed, accuracy and efficiency provided by the Microsart® Mycoplasma, Bacteria and Fungi qPCR kits provides peace of mind during your cell culture processes. These comprehensively validated kits are highly sensitive and offer broad range microbial detection, consisting of an efficient DNA isolation protocol, and followed by a real-time PCR assay using the Microsart® ATMP Bacteria/Fungi/Mycoplasma kit.

  • Accurate results in just three hours
  • Detects a broad range of bacteria and fungi
  • Highest level of qPCR specificity

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Sartorius Advanced Cell Model Solutions for Disease Modeling

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Improve clinical translation

Sophisticated solutions such as Incucyte® Live-Cell Analysis and CellCelector Automated Cell Selection and Retrieval Platform enable better decision making through monitoring and characterization of key parameters of advanced cell cultures such as size, count and morphology. 

Save precious sample material

Gain more information from one experiment using the Incucyte® Live-Cell Analysis System for dynamic monitoring in combination with the Incucyte® Organoid Analysis Software Module to simplify and facilitate temporal assessment of organoid growth or death. 

Streamlined protocols for 3D Models

The iQue® Advanced High-Throughput Flow Cytometer with associated suite of T cell characterization reagent kits, and validated spheroid washing and dissociation protocols provide an end-to-end solution for the evaluation of T cell response in advanced 3D tumor models.

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Resources for Advanced Cell Models

Organoid Analysis Guide
eBook

Organoid Analysis Guide

In order to effectively use organoid models specific and reliable culture and analysis methods are required; learn how.

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Quantitative Live-Cell Analysis for Optimization of Culture Conditions and Evaluation of Cell Health
Application Note

Quantitative Live-Cell Analysis for iPSC Optimization

Quantitative live-cell analysis for optimization of culture conditions and evaluation of cell health in human iPSC-derived neurons.

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Label-Free, Real-Time Live-Cell Assays for 3D Organoids Embedded in Matrigel®
Application Note

Real-Time Live-Cell Assays for 3D Organoids

Learn how the Incucyte® Live-Cell Analysis System with Organoid Analysis Software Module can simplify assessment of organoid growth or death.

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Frequently Asked Questions

The Incucyte® platform can be used to quantify multiple key metrics kinetically and objectively (i.e., formation, growth and morphology) to assess organoid expansion and growth efficiency during extended culturing periods.

Morphology metrics are cell-type specific and include indicators of maturation such as budding and accumulation of debris within the organoid lumen for intestinal organoids.

Real-time tracking of changes in organoid eccentricity (object roundness) and darkness (object brightness) with the Incucyte® platform enables rapid, unbiased assessment of optimal culture passage periods.

Many established methods require that cells need to be labeled with fluorescent probes for analysis; however, labeling can perturb biological responses and is often unsuitable for primary tissue. 

With the Incucyte® Live-Cell Analysis System (used in combination with the Incucyte® Organoid Analysis Software Module) you can avoid the need to manipulate the underlying biology (e.g., through recombinant expression of a fluorescent protein such as GFP), label-free live-cell imaging both saves researchers time and avoids any perturbation caused by fluorescent probes.

When using cancer cell lines traditionally grown in 2D to create 3D spheroids using either scaffold-free or scaffold-based techniques, we are using the recommended media for the 2D culture. When culturing 3D organoids, we are using specialized organoid media.

One method commonly adopted model is of single tumor spheroid invasion into an extracellular matrix such as Matrigel or Collagen. Here a single spheroid is created in a low adhesion plate, followed by direct addition of Matrigel into the same well, which provides a semi-solid matrix for the tumor cells to invade.

The advantages of this method include precise control of the spheroid size allowing you to mimic the defined micro-regions of metastatic solid tumors and since the invasion assay is performed in situ there is no need to transfer the spheroid to a second plate which is common with other methodologies. This method is also highly compatible with automated live-cell analysis. 

The CellCelector uses cooled deck tray can maintain hydrogel temperature at ~0 °C, thus preventing any polymerization before the organoid structure is deposited. 

It includes automatic morphology measurements and gating and identifies desirable organoids based on a range of morphological parameters, including area, diameter, sphericity, and the presence of neighboring organoids.

Additionally, due to the very gentle transfer the organoids retain their morphology and structure. 

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