Sartorius eBook - 30

THE FUTURE OF DRUG DISCOVERY: LIVE 3D CELL ANALYSIS

can occur over time12. Therefore, QC should be

are continually stirred to form aggregates, and

performed at every stage to monitor the health

agitation-free methods, where gravity-forced

and function of neuronal cell lines.

aggregation occurs13. These processes promote
cell-to-cell adhesion and typically induce the self-

4. Live Cell Monitoring of Complex

assembly of multicellular spheroids that secrete

Assays:3D Spheroid Formation

their own extracellular matrix components (ECM).

The creation of three-dimensional (3D) cell

Scaffold-based spheroid systems, contrarily,

culture systems is a key focus in drug discovery

mimic the native ECM and replicate the

and tissue engineering. These techniques

microenvironment of specific tissues in order to

demonstrate increased complexity and

promote cell growth and migration14. These can

translatability to the in vivo environment-and

be created from many different synthetic and

what's more, cells in 3D structures are often

biological materials, from microcarrier beads

better at maintaining differential and clonal

and micropatterned plates, to sophisticated

expansion after long-term passaging than cells

microfluidics systems. Natural ECM gels, such as

in 2D structures. In providing more relevant

Matrigel® and polymeric structures like hydrogels

and translational in vitro models compared to

provide structural matrices15, whereas collagen,

their 2D counterparts, 3D cell cultures are now

laminin, or alginate can be used to construct

gaining traction. Advanced 3D structures, such as

prefabricated scaffolds consisting of a network of

spheroids and organoids, better replicate the in

fibers through which cells can easily migrate16.

vivo cellular environment and microarchitecture

ECM and stromal cell scaffolds more closely

of organs, and show huge potential, particularly

recapitulate the tumor microenvironment, and

in the fields of oncology and hepatotoxicity.

therefore, increase the predictive accuracy of

Depending on the type of cell and tissue
origin, and the aims of the assay, 3D cultures

spheroids for testing compounds.
Despite their contribution to many areas of

can use scaffold-free or scaffold-based systems.

research, the formation of reproducible and

Scaffold-free methods that generate spheroids

uniform 3D spheroids assays is technically

are proving popular by offering a relatively easy

challenging and may follow unpredictable

route to 3D cell culture that can be applied to

time-courses. Additionally, 3D models exhibit

a broad range of cell types. Broadly speaking,

significantly more complex morphology and

scaffold-free techniques fall into agitation-

function than 2D cultured cells, not to mention

based methods, where cells in suspensions

their diversity in architecture, highlighting the

30 |

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Sartorius eBook

Table of Contents for the Digital Edition of Sartorius eBook

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