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How to Choose the Right 3D Cell Culture Platform

There are two main approaches for establishing 3D cell culture systems – spheroids and organoids, so which should you choose? They are often used interchangeably due to their similarities. However, their differences will generally make one more suitable than the other. A choice of either a scaffold-based or scaffold-free system, each with its pros and cons, will depend on the 3D cell culture required.

Understanding their specific qualities will help decisions over which is better suited to the task. To aid this decision, with the help of Manchester Biogel, we look deeper into what they are, the features, benefits and differences, and how they both work.

An organoid grown most commonly on a scaffold-based system using natural or synthetic materials or a spheroid grown on scaffold-free systems relying on encouraging self-aggregation of cells on specialised culture plates are the primary approaches used.

Spheroids Vs Organoids

Whilst a scaffold can be used for spheroid formation, often a scaffold-free, 3D cell culture technique is used. Spheroids are fairly simple structures and are usually generated by cell aggregates or a single cell type. It floats freely in ULA (or ultra-low attachment) plates.

Organoids, however, are scaffold-based 3D cultures of an organ or tissue-specific cell type, and are more complex than spheroids. They generally originate from stem cells and progenitors that have been embedded in a hydrogel matrix (ECM).

An organoid will better mimic in vivo environments in comparison with traditional 2D culture systems or spheroids. For this reason, organoids are now more widely used in diseased tissue modelling and for drug toxicity, efficacy testing and drug response, especially in drug development and targeting.

Two Major Differences

Spheroids develop primarily via cell-to-cell adhesion, whilst organoid formation is through internal development processes.

Organoids are an in vitro model of human development and disease to display accurate microanatomy (mini-organs). Organoids derive and maintain a population of stem cells during in vitro culture to guarantee long term viability by using a basement membrane matrix. When organoids are passaged, they retain genetic features of the original organ over multi-generations. Whichever stem cells are used to establish organoids, the link uses stem cells capable of self-renewal and the ability to differentiate into multiple lineages in vitro.

Spheroids, however, consist of cell aggregates generated from either a single cell type or a mixture of multicellular cells. When multicellular combinations of different origins are used, an organotypic culture results, suggesting a multicellular aspect of tissue in vivo. They are easy to generate, cost-effective, but challenging to maintain long-term cultures.

Spheroids better recapitulate in vivo features than 2D cultures and provide a simple platform for evaluating CSC characteristics and targeting drugs. However, for those seeking to move to a 3D system that will capture more physiological original organ relevance, a more suitable choice would be organoids. Organoids can be expanded to disease-specific organs and derive living biobanks of tissue. Their use is increasing for predicting patient drug response and within drug discovery applications. Organoids are better able to mimic the in vivo environment when compared to spheroids and traditional culture systems. However, they may require fine-tuned environments.

Platforms for Growth

Animal-free synthetic peptide hydrogels (PeptiGels ®) provide highly suitable platforms to support the growth of both spheroids and organoids, including liver, gut, cardiac, kidney and cancer. PeptiGels ® are reproducible, chemically defined peptide hydrogels, which provide a cost-effective, stable and entirely tenable platform for cell growth.

Such hydrogels are optimised for 3D bioprinting and biologically relevant to mimic the cell microenvironment with no preparation. They can be added to cells at room temperature and still achieve results, removing an element of the fine-tuning required. Bespoke design services allow optimal synthetic hydrogels to meet the growing need for 3D and 2D cell culture, tissue regeneration, drug discovery and bioprinting to support both organoid and spheroid growth.

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