Omar Khalid
Tissue culture treatment is a process that prepares plastic surfaces for cell adhesion and growth. This process is necessary because untreated plastic surfaces, such as polystyrene, are hydrophobic and not suitable for cell attachment. Tissue culture treatment involves exposing the plastic to plasma gas, which modifies the surface chemistry to make it more hydrophilic and suitable for cells to anchor and grow. This treatment has been used to develop cell culture vessels like dishes, flasks, and plates, which are used for various applications, including the industrial-scale production of vaccines and antibodies. The properties of these treated surfaces can be further optimized through coatings of peptides, proteins, or polysaccharides, enhancing cell adhesion and growth for specific cell types.
| Characteristics | Values |
|---|---|
| Plastic types used for cell culture | Polyethylene terephthalate (PET), high- and low-density polyethylene (PE), polyvinyl chloride (PVC), polypropylene (PP), and polystyrene (PS) |
| Most frequently used plastic | Polystyrene (PS) |
| Properties of PS surfaces can be optimized by | Coating with peptides (e.g., poly-D-lysine or PDL), proteins (e.g., collagen), or polysaccharides |
| PS surface treatment | Plasma treatment, liquid surface deposition, energetic plasma activation, functionalization methods |
| Plasma treatment | Exposing a PS surface to a plasma gas, which modifies the polymer chain and leaves oxygen-containing functional groups |
| Plasma-treated PS characteristics | Hydrophilic, negatively charged, suitable for cell adhesion, and enables subsequent coatings |
| Coating options | Poly-lysine, ECM proteins such as collagen |
| PS advantages | Low cost, inert chemistry, hydrophobic (ideal for suspension cell culture), optical clarity, ease of manufacture |
| PS disadvantages | Poor cell adhesion and growth in its native form |
| PS applications | Two-dimensional (2D) tissue culture, three-dimensional (3D) culture models through casting, electrospinning, 3D printing |
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Plasma treatment
The plasma treatment replaces the hydrophobic phenyl groups present on untreated polystyrene surfaces with hydrophilic functional groups, such as carbonyl, hydroxyl, or amine groups. These functional groups are determined by the process gas used in the plasma treatment. The resulting surface carries a net negative charge due to the presence of oxygen-containing functional groups.
The introduction of these hydrophilic functional groups improves the biocompatibility and seeding efficiency of the polystyrene surface. It increases the nonspecific adsorption of cell media constituents and enables subsequent coatings that further promote cell adhesion. This treatment process is crucial for culturing anchorage-dependent cells, which require a surface that allows cell adhesion and spreading.
Additionally, plasma treatment can be used to optimize the contact angle of the surface for specific applications. A customized contact angle can enhance the application of biologically relevant coatings. Researchers can also use spin coating techniques with plasma-treated polystyrene to control the substrate's topography while increasing the surface area and permeability.
Overall, plasma treatment of plastic for tissue culture is a widely adopted method due to its ability to improve cell adhesion, viability, and biocompatibility. It plays a crucial role in enhancing the biological affinity of the treated plastic, making it a preferred choice for cell culture applications.
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Coating the surface
The tissue culture treatment process involves exposing the polystyrene surface to a plasma gas, which replaces the hydrophobic phenyl groups with hydrophilic carbonyl, hydroxyl, or amine-containing functional groups. This process modifies the polymer chain, leaving behind oxygen-containing functional groups that increase cell attachment. The resulting surface carries a net negative charge, which can be further enhanced by using poly-lysine, a synthetic positively-charged polymer.
The properties of PS surfaces can be further optimized by coating them with peptides (e.g., poly-D-lysine or PDL), proteins (e.g., collagen), or polysaccharides. PDL is a chemically synthesized, polycationic, extracellular matrix (ECM) that can help mediate the negative charges of both the cell membrane and surface, thereby facilitating cell adhesion. ECM proteins such as collagen provide an attachment framework for the adhesion and growth of certain cell types that have difficulties growing on regular TC-treated surfaces.
In addition to the standard plasma treatment, there are other treatment methods available to alter PS surfaces and improve cell growth. These include liquid surface deposition, energetic plasma activation, and emerging functionalization methods that transform the surface chemistry. These methods allow for the incorporation of moieties containing oxygen and nitrogen, presenting a surface chemistry for cells to anchor and grow.
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Using polystyrene
Polystyrene (PS) is a commonly used plastic in labs today for cell culture. Its low cost, inert chemistry, optical clarity, ease of manufacture, and compatibility with many cell strains make it an optimal choice for a disposable culture surface.
However, untreated polystyrene surfaces are composed mainly of hydrophobic phenyl groups, which are not found naturally in the body and are detrimental to cell anchorage. To overcome this, polystyrene must be subjected to a tissue culture treatment process, also known as plasma treatment, to render the plastic suitable for cell attachment. This process involves exposing a polystyrene microplate to a plasma gas, which modifies the hydrophobic plastic surface to make it more hydrophilic. The resulting surface carries a net negative charge due to the presence of oxygen-containing functional groups such as hydroxyl, carboxyl, or amine groups. This treatment process increases cell attachment and adhesion, which is necessary for the growth of most cells derived from vertebrates.
The properties of PS surfaces can be further optimized by coating the surface with peptides (e.g., poly-D-lysine or PDL), proteins (e.g., collagen), or polysaccharides. PDL can help mediate the negative charges of both the cell membrane and surface, thereby facilitating cell adhesion to TC-treated plastic. ECM proteins such as collagen provide an attachment framework for the adhesion and growth of certain cell types that have difficulties growing on the regular TC-treated surface.
To transition PS to specialized 3D culture surfaces, methods such as casting, electrospinning, 3D printing, and microcarrier approaches can be utilized. Additionally, simulated body fluid (SBF) and protein adsorption layers can be used to functionalize PS plates with hydroxyapatite (HAp), promoting the adhesion and growth of mesenchymal stem cells.
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Cell adhesion
Polystyrene (PS) is the most frequently used plastic in labs today for 2D cell culture. This is due to its low cost and inert chemistry, making it an optimal choice for a disposable culture surface. However, in its pure form, PS is hydrophobic, which is not suitable for cell adhesion and growth in vitro.
To improve cell adhesion, the polystyrene surface must undergo a tissue culture treatment process. This involves exposing the PS surface to a plasma gas, which modifies the hydrophobic plastic surface to make it more hydrophilic. The plasma treatment introduces bioactive, hydrophilic functional groups to the cell culture materials, improving cell adhesion and cell viability. The resulting surface carries a net negative charge due to the presence of oxygen-containing functional groups such as hydroxyl and carboxyl, which leads to increased cell attachment.
The properties of PS surfaces can be further optimized by coating the surface with peptides, proteins, or polysaccharides. For example, poly-D-lysine (PDL) is a chemically synthesized, polycationic, extracellular matrix (ECM) that mediates the negative charges of both the cell membrane and surface, facilitating cell adhesion to TC-treated plastic. Similarly, ECM proteins such as collagen provide an attachment framework for the adhesion and growth of certain cell types that have difficulties growing on regular TC-treated surfaces.
Plasma treatment is also used to improve the cell adhesion properties of other materials, such as Polyethylene Terephthalate (PET), which is commonly used for ligament or vasculature grafts.
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3D printing
Three-dimensional (3D) printing technology has become an increasingly versatile experimental platform in the life sciences. It has been used to create tissue culture dishes, chambers, and bioreactors. 3D printing allows for the fabrication of complex structures and geometries that would otherwise be difficult to create using conventional techniques.
To create tissue culture-treated plastic using 3D printing, several steps are involved. First, a 3D model of the desired structure is designed using computer-aided design (CAD) software. This model is then sliced into layers, and a g-code is generated, which contains the instructions for the 3D printer. The g-code is then input into the 3D printer, which prints the plastic structure layer by layer.
When printing tissue culture dishes, it is important to consider the printing material and the printer hardware. Biocompatible and food-grade materials such as polylactic acid (PLA), polycarbonate (PC), and nylon can be used for 3D printing tissue culture dishes. However, the choice of material depends on the specific application, as some materials may be cytotoxic or incompatible with certain cell types.
The printer hardware should also be carefully selected to ensure accurate and precise printing. Modifications may be necessary to adapt the printer to printing tissue culture dishes. For example, the fan shroud may need to be adjusted to increase airflow, and the heater block may need to be replaced to fit the dimensions of the tissue culture dish.
In addition, surface treatment techniques such as plasma treatment or functionalization methods may be applied to the printed plastic to improve cell adhesion and growth. This can involve altering the surface chemistry or topography of the plastic to better facilitate cell culture.
Overall, 3D printing offers a cost-effective and versatile method for creating tissue culture-treated plastic. With the right materials, hardware, and surface treatments, it is possible to fabricate custom tissue culture dishes, chambers, and bioreactors that can enhance cell growth and provide a more realistic environment for cells.
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Frequently asked questions
Tissue culture-treated plastic is plastic that has been treated to allow cell adhesion and spreading. This treatment process involves exposing a polystyrene microplate to a plasma gas, which modifies the hydrophobic plastic surface to make it more hydrophilic.
Untreated polystyrene surfaces are not suitable for cell attachment due to their surface chemistry. Tissue culture treatment modifies the surface chemistry, allowing cells to anchor and grow.
Polystyrene (PS) is the most frequently used plastic in labs for tissue culture due to its low cost and inert chemistry.
The properties of polystyrene surfaces can be optimized by coating the surface with peptides (e.g., poly-D-lysine), proteins (e.g., collagen), or polysaccharides. These coatings help facilitate cell adhesion and provide an attachment framework for certain cell types.
