Spheroids Culture

Spheroids Culture

Spheres are dense 3D cell aggregates that exhibit extensive intercellular adhesion, usually retain their endogenous extracellular matrix, and have characteristics very similar to their tissue counterparts in vivo. They can better simulate tissue characteristics in vivo and have excellent differentiation ability compared with 2D culture system. These characteristics make them very suitable for drug development and tissue research, as well as the study of basic biological principles.

Cortical spheroids confocal projectionFigure 1.Cortical spheroids confocal projection (Dingle Y T L, et al. 2015).

Spheroids Culture

There is a growing trend to develop screening analyses using three-dimensional (3D) cell culture. A common method is to culture the cells in an extracellular matrix to allow greater interaction with neighboring cells and form more complex structures. CD BioSciences can culture different spheres through scaffold-based technologies and scaffold-free technologies, and monitor the growth of living cells with fluorescent nuclear staining and label-free imaging techniques.

Schemes of technical methods.Figure 2.Schemes of technical methods. (a) Pellet Culture, (b) Liquid Overlay, (c) Hanging Drop, (d) Spinner Culture, (e) Rotating Wall Vessel, (f) Microfluidics, (g) Magnetic Levitation (Ryu N E, et al. 2019)

Spheroids Culture Technologies

Technologies Method Properties
Scaffold-based technologies Pellet culture Use centrifugal force to concentrate cells
Liquid overlay Use non-adhesive materials to inhibit cell attachment
Hanging drop Use surface tension and gravitational force
Spinner culture Use convectional force by stirring bar
Rotating wall vessel Use constant circular rotation of vessel
Microfluidics Use microfluid flow and materials permeable to soluble factors
Magnetic levitation Use magnetic force to levitate cells
Scaffold-free technologies Hydrogel  Entrap cells during culture and can deliver cells as injectable form.
Provide an environment similar to extracellular matrix and improve viability, stemness and angiogenetic capacity of stem cells.
Biofilms Increase stemness, differentiation potential, adhesion and proliferation of stem cells
Particles Control mechanotransductional mechanisms inside the spheroid and improve viability and proliferation

Spheroids Culture Workflow

The following is the general process of spheroids culture:

Step 1


Choose cell lines that allow you to closely mimic the in vivo state and generate more relevant data.

Step 2


Selecting the right culture matrix is an important first step in developing a successful culture system for spheroids.

Step 3


Your 3D culture system can be modified and developed to suit specialized cell types like stem cells or cancer cell lines.

Step 4

Monitor & analyze

Before conducting experiments, it is necessary to monitor spheroids and describe their characteristics.

Step 5

Characterize and assay

Molecular characterization and applications of 3D spheroid models.


Spheroids size distribution
Spheroids volume
Number of cells

Cell viability
Mitochondrial function
Expression of specific markers
Other relevant data

Our Advantages

  • Confocal imaging and 3D image analysis
  • Provide three-dimensional information of cells
  • Experienced scientists provide experimental consultation
  • Reasonable price and short turnaround time

CD BioSciences has technicians with rich working experience in the field of 3D cell culture. We can design a personalized 3D cell culture program for you to obtain spheroids that meet your needs, and we also provide you with spheroids quantitative analysis reports. If you have any needs, please feel free to contact us.

  1. Ryu N E, Lee S H, Park H. Spheroid culture system methods and applications for mesenchymal stem cells[J]. Cells, 2019, 8(12): 1620.
  2. Dingle Y T L, Boutin M E, Chirila A M, et al. Three-dimensional neural spheroid culture: an in vitro model for cortical studies[J]. Tissue Engineering Part C: Methods, 2015, 21(12): 1274-1283.

*If your organization requires the signing of a confidentiality agreement, please contact us by email.

Please note: Our services can only be used for research purposes. Do not use in diagnostic or therapeutic procedures!

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