3D Cell Culture Workflow Tools

Organoids are a collection of organ-specific cell types that exhibit in vivo-like organ anatomy, functionality, and phenotypes. They can be derived from patient tissues (patient-derived organoids; PDOs) or induced pluripotent stem cells (iPSCs).

Organoids have been used to study organ-specific disorders and diseases including related cancers, inflammatory bowel disease, and cystic fibrosis. They are also used to study viral respiratory disease, drug transport, drug permeability, ADME/Tox, and organogenesis. Organoids are typically cultured using hydrogels.

We provide organoids derived from patient tissues, induced pluripotent stem cells, and answers to frequently asked questions about organoid culture:

Organoid Culture Frequently Asked Questions

Hydrogels are a cellular tool that researchers use to mimic the natural cellular environment in vitro. As a water-swollen network of polymers, hydrogels remain in the liquid state at 4˚C. This allows researchers to ebbed cells before the gel forms at incubation temperatures of 37˚C. The cells then become encapsulated in the gel and can be cultured in an environment that is physiologically relevant.

Need to choose a hydrogel? Learn about the key considerations in hydrogel selection for 3D cell culture applications, including:

• Natural vs. synthetic hydrogels
• Biological relevance
• Reactivity
• Ease of use
• Reproducibility
• Customization
• Formats including powders, liquids, kits, and pre-cast microplates

Formed by the self-aggregation of cells, spheroid cultures are excellent models of tumor-related disorders and diseases such as cancer. Because spheroids have surface cells and buried cells, these models are commonly used to tumor formation, tumor hypoxia, and drug penetration. 

Spheroids are easily amenable to both high throughput and co-culture applications. They can be grown using scaffold-free technologies such as ultra-low attachment (ULA) plates or scaffold-based technologies such as hydrogels.

• Cancer Stem Cell Tumorsphere Formation Protocol
• Tumor Spheroid Formation Assay

Organotypic culture systems (OCSs) culture organs in vitro that were collected from an organism. These models preserve normal organ architecture and interactions while supporting the formation of nearly normal stratified epidermis cells. They can be used to mimic complex pathologies of the human skin, airway, intestine, liver, heart, and neurological tissues.

Organotypic models are commonly used to study a variety of related cancers, aging, autoimmune disorders, infection, tissue microenvironment, and tissue development. These systems are typically cultured using scaffold-based technologies such as porous membrane-based cell culture inserts.

• Generating Organotypic 3D Skin Cultures as an In Vitro Model of the Human Epidermis
• Using Immortalized 16HBE14o- Human Bronchial Epithelial Cell Lines to Model Respiratory Lung Diseases
• Cell Migration Assay Guidance using Millicell^® Hanging Cell Culture Inserts

Culturing organoids, spheroids, and other 3D models often require specific tools to support the establishment of the cultures. Researchers could use separate scaffolds for their cultures, or they can culture cells directly into 3D cell culture plates. Plates with ultra-low attachment (ULA) coatings, or that contain additional microwells promote organoid, tumoroid, and spheroid formation in a controlled 3D environment.

Millicell^® Ultra-low Attachment (ULA) Plates

Millicell^®ULA plates promote scaffold free self-assembly of spheroids using a stable, non-binding polymer. The plates contain a ultra-hydrophilic polymer that facilitates spontaneous and natural spheroid formation. Millicell^®ULA plates have high optical clarity, making them suitable for both brightfield imaging and confocal microscopy.

We cultured multiple spheroid forming cells using Millicell^® ULA plates. Using the plates, we developed quantitative measurements of spheroid formation by calculating spheroid circularity and roundness.

Millicell^® Microwell Plates

Millicell^® Microwell plates contain an array of ultra-dense, U-bottom microwells (W) and hydrogel. The addition of microwells promote reproducible organoid culture for high throughput and screening applications. The microwells also allow researchers to generate up to hundreds of homogeneous organoids in a single well. As the organoids form in a single focal plane, Millicell^® Microwell plates support organoid seeding, imaging, and analysis in the plate.

Millicell^® Microwell plates are ideal for multiple applications, including blood brain barrier penetration assays and T cell killing assays.