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Mode of 3D Cell Culture

What is 3D Cell Culture? Classification of 3D Cell Culture?

In vivo, cells exist in three-dimensional aggregate/mass, and interface with the other cell or extra cellular matrix (ECM). In conventional in vitro cell culture method which is easy and has a long history, the cells attach to culture dish and form monolayer. This method have achieved many knowledge in biology. However, it is reported many times that the cells cultured in 2D method are far different from the cells in vivo. The excessive activation of proliferation signal caused by cells attachment to the plastic dish and poor (two-dimensional) interface with the other cells are the main reasons.

Recently, it is well known that cells grown under 3D cell culture technique more closely mimic natural tissues and organs microenvironment than cells grown in 2D. In 3D cell culture, cells form multi-cellular clusters (spheroids) that are driven by cell-cell and cell-matrix contracts, rather than cell-plastic contracts. In this environment, cells can exert forces on one another and can move and migrate as they do in vivo.

3D cell culture methods fall into two major groups. One is culturing in hydrogel sponge, e.g. ECM and agarose (i.e. scaffold type). Another is culturing in the low attachment dish (i.e. scaffold-free type). These methods have merit and demerit, and these are used according to the situation demand.

Scaffold Type Scaffold-Free Type
Pros
  • ・Has many past records and continuousness in research (standard method for 3D culture)
  • ・Cells cultured in this method mimic in vivo atmosphere, and have a high proliferative character
  • ・Suitable for construction of structural tissue
  • ・Many Plate-type products, which can be handled like conventional 2D culture plate, are available
  • ・Plastic products with high uniformity
  • ・Suitable for HTS
Cons
  • ・Scaffold made with biological product has problem in the stability and reproducibility.
  • ・Cumbersome procedure (coating and/or icing).
  • ・Not suitable for high throughput observation, because spheroids exist in different depth in the scaffold.
  • ・Cells proliferation under this condition is low, because proliferation signal from cell adhesion molecule is poor.
  • ・Cells form spheroid engulfing dead cells, and/or a lot of cells die in the spheroid because of low nutrients and low oxygen .

Scaffold-based 3D cell culture biological products are usually using for basic research like cancer, regenerative medical and ADME/Tox. However, it is recognized that this method is not suitable for experiments/tests, which need a high reproducibility and throughput, such as drug screening method, because of lot-to-lot variation and cumbersome procedure.

Our products, NanoCulture Plate and NanoCulture Dish, is the scaffold type 3D culture system without any hydrogels or chemical pre-treatments. It is a nice combo of both type. It can be handled like conventional 2D culture plate but cells cultured on NCP have high proliferative character, high uniformity and suitable for HTS.

Scaffold type

Biological matrix
MatrigelTM (Corning)
Collagen
Soft agar
NanoCulture Plate
Artificial scaffold
Fiber
Sponge/mesh

Scaffold-free type

Hanging drop
Low attachment plate
Round-bottom
Flat-bottom
Low attachment coating
Poly-HEMA
Rotary culture