When it comes to cell culture, selecting the right culture vessels is crucial for optimal cell growth and accurate experimental results, along with the use of suitable culture reagents and ideal conditions. NEST offers a variety of surfaces with different treatments to ensure the best outcomes for cell culture with different cell lines. Let's explore the different options of cell culture surfaces:

1. TC-treated surfaces for adherent cells:

NEST utilizes TC treatment, a surface modification technique that enhances cell adhesion and improves culture efficiency. Many of our cell culture consumables undergo this treatment, facilitating better adherence and increased efficiency in cell culture.

2. Untreated surfaces for suspension cells:

For suspension cell culture, NEST provides high-quality, transparent polystyrene surfaces that are hydrophobic. These surfaces are ideal for the growth of suspension cells and are suitable for various biochemical assays.

3. Ultra low-attachment surfaces for 3D cell culture:

NEST also offers ultra low-attachment surfaces with a unique hydrogel coating. This coating prevents both specific and non-specific attachment, making it suitable for the 3D cell culture process. These surfaces are non-toxic, biologically inert, and non-degradable, and are utilized in a variety of culture vessels.

By offering a diverse range of cell culture surfaces, NEST ensures that researchers can choose the most suitable surface for their specific cell lines and experimental requirements.

Why is ultra low-attachment surfaces essential for 3D cell culture?

In traditional 2D cell culture, cells only adhere to one side of the culture surface, causing gradual flattening over time. This can lead to abnormal cell division and the loss of their differentiated phenotype in certain cases.

In 3D cell culture techniques, the goal is to replicate the intricate cell-to-cell and cell-to-matrix interactions that occur in the body. By allowing cells to attach to the entire surface of the culture vessel, ultra low-attachment surfaces enable the formation of 3D cell spheroids. These spheroids mimic the natural cellular architecture and facilitate the exchange of nutrients and growth signals among the cells.

By keeping cells in suspension, ultra low-attachment surfaces prevent the flattening process and abnormal cell division that often occur on flat culture surfaces in vitro. This effectively addresses the issue of cells losing their differentiated phenotype when cultured in 2D. By maintaining a more physiological cellular environment, 3D cell culture systems utilizing ultra low-attachment surfaces provide researchers with the opportunity to study cell behavior and function in a more representative manner.

NEST 3D Cell Culture Collection compasses...

1)  Ultra-Low Attachment Cell C ulture Plates : the surface  has extremely low surface tension , effectively preventing the attachment and reducing the interaction between cells and the surfaces as well .  The unique features allows specific  types of cells to maintain better morphology and vitality during the cultivation process.

Features

1.  Promotes the formation of 3D multicellular spheroids.

2.  Maintains cells in a suspended, non-adherent state.

3.  Prevents differentiation mediated by stem cell attachment.

4.  Inhibits anchorage-dependent cell division.

5.  Reduces the binding of low-attachment proteins and serum proteins to the substrate.

Application

1.  Cultivation  of 3D multicellular spheroids, such as embryoid bodies and tumor spheroids.

2.  Cultivation  of neural spheres in Neurobiology

3.  Cultivation  of embryoid bodies in Stem Cell biology

4.  Studies on the activation and deactivation mechanisms of lymphocytes, monocytes, macrophages, and other phagocytic cells.

5.  Suspension-based cell screening

6.  Cultivation of cells, tissues, or organ fragments.

7.  Cultivation  of multicellular spheroids for t umor research , etc.  

2)  GelNest ™  Matrix: it is derived from mouse tumor tissue and contains extracellular matrix components such as laminin, type IV collagen, heparan sulfate proteoglycans, and more. These components support cell adhesion, differentiation, and proliferation, providing signals for these processes. Additionally, they simulate the characteristics of the basement membrane in the physiological environment, enhancing the success rate and effectiveness of cell culture.

In addition to the matrix components, GelNest ™  Matrix is rich in various growth factors. These growth factors promote cell differentiation, proliferation, and migration, mimicking cellular signaling pathways and interactions in the physiological environment. GelNest ™ Matrix has a wide range of applications, particularly in tissue engineering, cell culture, and research. It can be used for organoid culture, stem cell differetiation, angiogenesis, migration or invasion assays, and in vivo tumor studies.

How is the cultivation performance of NEST Ultra-low Attachment Surfaces?

To validate their performance, we launched the cultivation experiment with HUVEC cells and brain organ cells within our Ultra-low Attachment Culture Plates and further recorded the results.

HUVEC cell culture

Conclusion: As seen in the graph, cells cultured in the plates can maintain a desirable suspension state. Compared to TC-treated products, the attachment on the Ultra-low Attachment Culture Plates is reduced by 98%.

NEST Ultra-low Attachment Plate Ordinary Cell Culture Plate

Brain organ cell culture

Brain organ cells were seeded in NEST Ultra-low adhesion cell culture plates, and the growth of brain organ cells at different stages was observed and recorded.

Day 2-Day 4:

Formation of EB (embryoid bodies): Seed the cells in the plates and culture until round and smooth edges appear.

Day 6-Day 7:

Induction of differentiation: transfer the EBs to the plates and observe them with semi-transparent edges on optical inspection.

Day 9-Day 10:

Amplification: embed the EBs in GelNest ™ Matrix Gel and continue culturing them in the plates. The EBs will start budding (amplification of neuroepithelial cells) on the surface.

Conclusion : During the culture process within the plates, the cells did not attach to the surface, avoiding experimental failure. Moreover, the cells remained stable for a long time, successfully enabling their growth, differentiation, and amplification. NEST Ultra-low Attachment Culture Plates are an excellent choice for brain organ culture experiments.

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