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.
Figure 1．Cortical spheroids confocal projection (Dingle Y T L, et al. 2015).
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.
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
|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:
Choose cell lines that allow you to closely mimic the in vivo state and generate more relevant data.
Selecting the right culture matrix is an important first step in developing a successful culture system for spheroids.
Your 3D culture system can be modified and developed to suit specialized cell types like stem cells or cancer cell lines.
Monitor & analyze
Before conducting experiments, it is necessary to monitor spheroids and describe their characteristics.
Characterize and assay
Molecular characterization and applications of 3D spheroid models.
Spheroids size distribution
Number of cells
Expression of specific markers
Other relevant data
- 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.
- Ryu N E, Lee S H, Park H. Spheroid culture system methods and applications for mesenchymal stem cells[J]. Cells, 2019, 8(12): 1620.
- 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.
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