3D cell culture enables cells to grow, proliferate and migrate in a three-dimensional space to form a three-dimensional cell-cell or cell carrier complex, so as to better simulate the growth environment of cells in vivo. 3D cell culture has the advantages of approaching all aspects of human tissue, such as structure, cell tissue, interaction between cells and cell matrix, and more physiologically related diffusion characteristics. Therefore, 3D models are used in drug screening and cancer research.

Figure 1．Floating spheroids, microtissues or cellular clusters.

Currently, 3D cell culture is divided into scaffold and scaffold free three-dimensional culture technology. 3D cells cultured on scaffolds are inexpensive and simple to operate. Scaffold materials include collagen and hydrogel. 3D cell culture without scaffolds is mainly carried out by physical methods, mainly including microcarriers, magnetic levitation, hanging drop plates, etc. These types of operations are complicated and costly.

Characteristics of Different 3D Cell Culture Methods

Type of 3D system	Description of cell culture	Advantages
Suspension cultures on non-adherent plates	Single cells are seeded on non-adherent plates with medium 3D structures can be observed after 3 days of culture	Simplicity, easiness and speed of conducting culture Bacterial plates or non-adherent culture plates can be used but only for some cell lines Cells can be easily extracted from the medium and used for further experiments
Cultures in concentrated medium or in gel-like substances	Single cells grow in medium containing substances with gelling properties: i) dissolved low-melting agarose with cell medium is poured on plate and incubated until solidifying to obtain the first, lower layer; the top layer consisting of agarose and the medium with single cells is added; ii) the cells are flooded in Matrigel (multiprotein hydrogel) 3D structures can be observed after 7 days of culture	Soft agar allows studying both the growth of a single cell regardless of attachment and the phenomenon of escape from anoikis Cells can be easily recovered for further analysis Cells have three-dimensional interactions with the local environment and form tissue-like structures Used to study the aggressiveness of the cells and their potential for metastasis
Cultures on scaffold	The cells can migrate among fibres and attach to the scaffold, made of biodegradable material such as silk, collagen, laminin, alginate, and fill the space among fibres, grow and divide	System is compatible with commercially available functional tests, as well as with DNA/RNA and protein isolation kits Easy to prepare for immunohistochemical analysis

3D Cell Culture and Imaging Analysis Workflow

Although imaging analysis of 3D cell models has become an extremely attractive research tool for understanding complex biology, 3D cell image acquisition and analysis methods are still important factors hindering its development. CD BioSciences can generate more predictive data by image acquisition and analysis of 3D cell models through high connotation imaging (HCI) system. The following is the general flow of 3D cell culture and imaging analysis.

Delivery

Our Advantages

• Confocal imaging and 3D image analysis
• Provide three-dimensional information of cells
• Provide various types of organoids
• 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 organoids and spheroids, and we also provide you with their imaging analysis reports. If you have any needs, please feel free to contact us.

*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!