Ethan Clarke
Plastic cell culture vessels are popular in research because they are disposable, optically clear, easy to mould, and can be sterilized by irradiation. However, they are also very hydrophobic, which makes it difficult for cells to attach to their surfaces. To improve cell attachment, the hydrophobic polystyrene surface can be modified to be more hydrophilic, allowing cell attachment proteins (vitronectin and fibronectin) found in the serum-containing culture medium to adhere and spread on the vessel bottom, providing a better surface for cells to attach. This process is not yet fully understood, and there is still a lot to learn about cell adhesion, proliferation, and apoptosis.
| Characteristics | Values |
|---|---|
| Cell Adhesion | Indirect |
| Cell Culture Plates | Coated with negatively charged groups |
| Plastic Culture Vessels | Serum and serum components can interfere with cell attachment |
| Plastic Plates | Cells don't always adhere |
| Plastic Surfaces | Hydrophobic, requiring modification to a hydrophilic surface for cell attachment |
| Cell Culture Surfaces | 2D nature of traditional polystyrene and glass surfaces may not be satisfactory for promoting in vitro cell functions |
| Cell Culture Surfaces | Ultra-Web® synthetic nanofiber surfaces offer cells a more in vivo-like 3-D fibrillar topography |
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What You'll Learn
- Plastic culture vessels are treated with negative charges, attracting proteins from the media
- Cell attachment proteins, like vitronectin and fibronectin, adhere to hydrophilic surfaces
- The hydrophobic nature of polystyrene makes it difficult for cells to attach
- Cells adhere to plastic as a substitute for the extracellular matrix
- Surface hydroxyl groups are important for the adhesion of baby hamster kidney cells
Plastic culture vessels are treated with negative charges, attracting proteins from the media
Plastic culture vessels are often manufactured from polystyrene, a hydrophobic polymer to which cells have difficulty attaching. To promote cell adhesion, the hydrophobic polystyrene surface is modified to a more hydrophilic surface. This allows cell attachment proteins (vitronectin and fibronectin) found in the serum-containing culture medium to adhere and spread on the vessel bottom, providing a better surface for cells to attach.
The net negative charge of the plastic culture vessel attracts proteins from the media, which act as 'adhesion factors'. These adhesion factors are recognised by molecules on the cell surface, which bind to them. This indirect form of cell adhesion is strong enough to keep dying or dead cells attached to the vessel surface.
The use of plastic culture vessels, such as microplates, flasks, dishes, and 96-well plates, became popular in the 1960s due to their optical clarity, ease of moulding, and ability to be sterilised by irradiation. However, the hydrophobic nature of polystyrene posed a challenge for cell adhesion.
To improve cell attachment, researchers have developed treatments such as the Corning® CellBIND® surface, which has shown enhanced attachment and yield of LNCaP cells compared to standard cell culture surfaces. More recently, Ultra-Web® synthetic nanofiber surfaces have been introduced, offering a more in vivo-like 3-D fibrillar topography that can improve cell performance and functionality.
While plastic culture vessels are commonly used, it is important to note that adhesion to plastic is not a perfect substitute for the extracellular matrix (ECM). For ECM-dependent processes like migration, adhesion, and invasion, it is recommended to coat plates with collagen or Matrigel to better mimic the in vivo environment.
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Cell attachment proteins, like vitronectin and fibronectin, adhere to hydrophilic surfaces
Cell attachment proteins, such as vitronectin and fibronectin, play a crucial role in facilitating cell adhesion to hydrophilic surfaces. These proteins are present in the serum-containing culture medium and have a strong affinity for such surfaces, promoting subsequent cell attachment.
Vitronectin, a glycoprotein composed of 459 amino acids, is a vital component of the extracellular matrix (ECM) and blood. It interacts with polysaccharides and proteoglycans, acting as a cell adhesion molecule. Vitronectin promotes cell migration, proliferation, differentiation, and the spreading of endothelial and neoplastic cells. Its ability to adhere to hydrophilic surfaces enhances cell attachment and growth, making it a key factor in cell culture optimization.
Fibronectin, on the other hand, is a large glycoprotein composed of two polypeptide chains joined by disulfide bonds. It is secreted by a wide variety of connective tissue cells, including fibroblasts, chondrocytes, Schwann cells, macrophages, intestinal epithelial cells, and hepatocytes. Fibronectin is a multifunctional protein that plays a significant role in cell adhesion, spreading, and cytoskeletal organization. It also regulates cellular morphology, cell migration, hemostasis, and wound repair. The presence of fibronectin in the culture medium contributes to its adherence to hydrophilic surfaces, providing a favourable environment for cell attachment.
The hydrophilic nature of a surface is a critical factor in promoting cell adhesion. Polystyrene, a commonly used material in cell culture vessels, is inherently hydrophobic, posing challenges for cell attachment. However, by modifying the polystyrene surface through treatments or coatings, it can be transformed into a hydrophilic surface. This modification increases the wettability of the surface, allowing cell attachment proteins like vitronectin and fibronectin to adhere and spread effectively.
The use of biological coatings, such as extracellular matrix proteins and synthetic polymers, can further enhance cell attachment and growth. These coatings provide a more consistent and stable surface for cells to attach and proliferate. Additionally, coatings like poly-D-lysine (PDL) create a positive charge on polystyrene, increasing the positively charged sites available for cell binding and improving attachment, especially in serum-free or low-serum conditions.
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The hydrophobic nature of polystyrene makes it difficult for cells to attach
Polystyrene is a long carbon-chain polymer with benzene rings attached to every other carbon. It is a hydrophobic polymer, meaning it is non-wettable, and this makes it difficult for cells to attach to it. This is a significant drawback of polystyrene for use in cell culture.
To improve cell attachment, the hydrophobic polystyrene surface must be modified to a more hydrophilic surface. This allows cell attachment proteins (vitronectin and fibronectin) found in the serum-containing culture medium to adhere and spread on the vessel bottom, providing a better surface for cells to attach. Corning® CellBIND® surfaces, for example, have been shown to significantly improve the attachment and yield of LNCaP cells.
In addition, 2D surfaces such as polystyrene and glass, even when coated with biological materials, are often not satisfactory for promoting in vitro cell functions such as transport or differentiation, which are done in a 3D environment in vivo. To address this, synthetic nanofiber surfaces have been introduced, offering cells a more in vivo-like 3D fibrillar topography that can improve their performance and functionality.
While plastic dishes are a very imperfect substitute for the extracellular matrix (ECM), they are still used in cell culture because they are disposable, have excellent optical clarity, are easy to mold, and can be sterilized by irradiation.
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Cells adhere to plastic as a substitute for the extracellular matrix
Plastic has been widely used as a material for cell culture vessels since the 1960s due to its optical clarity, ease of moulding, and ability to be sterilised by irradiation. However, cells have difficulty attaching to the hydrophobic surface of plastics like polystyrene. To address this, the plastic surface can be modified to be more hydrophilic, allowing cell attachment proteins (such as vitronectin and fibronectin) found in the serum-containing culture medium to adhere and spread, providing a better surface for cells to attach. This modification can be achieved through surface treatments such as coating the plastic with biological materials or using synthetic nanofiber surfaces that mimic the 3-D environment of the extracellular matrix (ECM).
The process of cell adhesion to plastic surfaces is complex and involves electrostatic reactions and interactions with the extracellular matrix. Individual cells do not directly "see" surfaces but instead sense the net negative charge that attracts proteins from the media, which act as 'adhesion factors'. These adhesion factors contain motifs that are recognised by molecules on the cell surface, leading to indirect cell adhesion. The specific mechanisms of cell adhesion can vary depending on the cell type, such as vascular endothelial cells, monocytes, or fibroblasts.
In the case of monocytes, scientists have encountered challenges with their adherence to plastic plates, which is a common method for isolating and separating different cell types. This non-adherence can interfere with the isolation process and requires alternative approaches, such as centrifugation or the use of specific detachment solutions.
While plastic surfaces have been modified to enhance cell adhesion, they still remain an imperfect substitute for the ECM. This discrepancy can affect cellular behaviours related to migration, adhesion, and invasion, which are ECM-dependent processes. To address this, researchers can coat their plates with collagen or Matrigel to more closely mimic the ECM environment and improve the accuracy of in vitro studies.
Overall, the adherence of cells to plastic surfaces involves a complex interplay between the surface properties, the culture medium, and the specific cellular mechanisms of adhesion. While plastic has been a convenient and widely adopted material for cell culture, ongoing research continues to enhance our understanding of cell adhesion and develop more optimal surfaces that better resemble the natural ECM environment.
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Surface hydroxyl groups are important for the adhesion of baby hamster kidney cells
Baby Hamster Kidney (BHK) cells are an important cell line derived from the Syrian golden hamster. They are used extensively in biopharmaceutical production and for the production of vaccines, particularly for animals.
The adhesion of cells to plastic surfaces is an important aspect of cell culture. In reality, individual cells do not 'see' surfaces. Instead, the net negative charge of the surface attracts proteins from the media, which act as 'adhesion factors'. The proteins on the cell surface then recognise and bind to these adhesion factors, leading to cell adhesion.
The importance of surface hydroxyl groups for the adhesion of BHK cells was demonstrated by the inhibition of adhesion when these groups were blocked. Hydroxyl (OH) functionalized molecules can act as a linker, reacting with proteins and accelerating cell growth, migration, and differentiation. Water vapour plasma treatment of polymer surfaces results in the formation of hydroxyl groups, which have been shown to have good adhesion properties with BHK cells.
In addition to hydroxyl groups, other factors can also influence cell adhesion. For example, the electrospinning technique is used to produce nanofibers that can enhance cell adhesion. Furthermore, specific receptor ligands can be coated onto surfaces to mediate cell attachment, and the stiffness of the surface can also play a role in cell adhesion.
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Frequently asked questions
Cells adhere to plastic as part of the scientific process of cell culture, which involves culturing cells in plastic culture vessels.
Cell culture vessels are often made from polystyrene, a long carbon chain polymer with benzene rings attached to every other carbon.
Polystyrene is used because it has excellent optical clarity, is easy to mould, and can be sterilised by irradiation. However, it is a hydrophobic polymer, so the surface must be modified to a more hydrophilic surface to allow for cell attachment.
The plastic surface is treated with a negative charge, which attracts proteins from the media. These proteins act as 'adhesion factors' and bind to the molecules on the cell surface.
