Most of the original plastic laboratory containers were made of polystyrene, a long polymeric carbon chain with benzene cores attached to every other carbon. Polystyrene was chosen for its excellent optical purity. It is easy to shape and relatively inexpensive. However, it also has one significant disadvantage: it is very hydrophobic (non-wetting), so it is difficult for cells to adhere to it. Fortunately, the surface of polystyrene can be easily modified by various chemical (sulfuric acid) or physical methods (corona discharge, gas plasma or irradiation). Using these methods, hydroxyl, ketone, aldehyde, carboxyl and amine groups can be attached polymorphically to the polymer. These groups change the surface characteristics, which make the uncharged hydrophobic surface more ionically hydrophilic. Polystyrene can also be modified by chemical reactions,
surfaces facilitating adhesion
- Corning CellBIND
- Tissue Culture Treated (TCT) – Standard polystyrene treated for tissue cultures
- Polystyrene coated with poly-D-lysine
- Ultra-Low Attachement – Polystyrene with a surface with low adhesion
- Untreated polystyrene – Untreated polystyrene
- Corning Osteo Assay
- Synthemax® II-SC substrate
|Corning CEllBIND – modified polystyrene||Hydrophilic and ionic (negatively charged)||Improves cell adhesion and binding to polystyrene|
|Standard polystyrene treated for tissue culture (TCT)||Hydrophilic and ionic (negatively charged)||It allows cell adhesion and binding to polystyrene|
|Untreated polystyrene||Hydrophobic||Significantly reduces the adhesion of most cells|
|Ultra-Low Attachement (Polystyrene with low adhesion coating)||Hydrophilic and nonionic||The hydrogel layer prevents the adhesion of most cells|
|Polystyrene coated with poly-D-lysine||Hydrophilic and ionic (positively charged)||Improves cell adhesion and binding to polystyrene|
|Synthemax® II-SC substrate||Surface treatment with synthetic peptide||It mimics the natural environment of cells with a peptide that promotes extracellular matrix cell adhesion|
|Corning Osteo Assay surface||Crystalline calcium phosphate||In vivo, it mimics the surface of bones in in vitro cell assays.|
Corning HYPER Flask ® Chambers
|Surfaces facilitating adhesion||Standard polystyrene treated for tissue cultures||X||X||X||X||X||X|
|Polystyrene coated with poly-D-lysine||X|
|Adhesion-preventing surfaces||Polystyrene with a low adhesion coating||X||X||X||X||X|
|Special surfaces||Synthemax® II-SC substrate||X||X|
|Corning Osteo Assay surface||X|
Surfaces facilitating adhesion
Patented microwave plasma surface that increases cell adhesion, reduces serum content and eliminates the need for scaling
useful for growing “difficult” cells, such as:
- primary culture
- transfected cells that overexpress proteins
- cells that normally require cornering
- increase adhesion, improve cell proliferation and improve yields
- improved cell adhesion under difficult conditions such as medium with reduced serum or serum-free medium, leading to higher cell yields (up to 60% higher yields at harvest)
This optimized surface treatment increases the oxygen content of the polymer surface, which leads to increased hydrophilicity and wettability and thus improves cell dispersion and adhesion. Importantly, the CellBIND® surface contains 68% higher oxygen levels than the standard tissue culture treated (TCT) surface and 105% more than competing amine surfaces.
The culture vessels are made of high quality polystyrene, which is shaped and then processed with energy and gas to create the surface roughness and hydrophilicity necessary for protein absorption and cell attachment. This treatment can be divided into two commercial types of treatment. Atmospheric plasma treatment in which a source of electrical energy is combined with atmospheric gases (standard corona discharge) to form a reactive plasma. The second group of treatments involves a vacuum plasma treatment in which an electric or radio frequency energy source is used in combination with a vacuum chamber and a pressurized gas containing oxygen or nitrogen / oxygen to form a reactive plasma. The CellBIND® surface treatment is a unique process that involves a high-energy microwave source, a vacuum chamber and a mixture of gases,
A. Untreated polystyrene chain
B. Corning CellBIND surface, treatment increased the number of oxygen-containing functional groups.
Plasma treatment of pure polystyrene with a highly reactive Corning CellBIND surface reduces the number of aromatic groups and increases the number of oxygen-containing functional groups.
Unlike biological coatings, Corning CellBIND does not require special handling or storage. Because the polymer is treated rather than coated, the surface is more consistent and stable.
- rapidly adapts cells to serum-free or serum-free conditions, thus eliminating the need for tedious, time-consuming, expensive, and poorly stable biological surfaces
- provides more uniform cell adhesion, even in cell lines that are difficult to adhere (especially transfected cells)
- reduces the premature separation of cells from confluent cultures, especially in cylinder flasks and cell assays
- increases cell survival after cryopreservation
- allows better recovery of primary cell isolates
- allows cell growth and protein production in conditions with a small amount of serum
- allows normal handling / automation
The Corning CellBIND surface is available for flasks, multiwell plates, Corning CellSTACK® culture chambers, cylindrical flasks, 96- and 384-well microtiter plates and petri dishes.
Recommendations for cell dissociation
Inoculation and harvesting of the culture should be performed by conventional methods. Both enzymatic and non-enzymatic dissociation solutions have been successfully used to remove cells from the CellBIND surface. These include: trypsin-EDTA, Accutase®, Versene®, Dispase® and citric saline. Some dissociating agents, such as Dispase or Versene, should be removed by centrifugation before reseeding the cells.
Increased adhesion of LNCaP cells to the surface of Corning® CellBIND®
Left: Recovery and growth of LNCaP adherent cells 24 hours after thawing. Average data obtained from 3 independent experiments. Right: Number of cells after 7 days of growth. Average data obtained from 3 independent experiments.
Tissue Culture Treated (TCT) – Standard polystyrene treated for tissue cultures
Standard polystyrene surface treated for tissue cultures – the classic and most used surface for growing cell cultures.
for routine cultivation
Standard culture vessels are surface modified either with a corona discharge (flask, dish, microplate) or with gas plasma (cylinders and culture tubes). These processes produce high-energy oxygen ions that integrate into the polystyrene chain (Fig. 1), so that the surface becomes hydrophilic and negatively charged when placed in the medium.
Corning offers a standard surface treated for tissue cultures on flasks, dishes, multiwell plates, CellSTACK culture chambers, cylindrical flasks and culture flasks.
Polystyrene coated with poly-D-lysine
- 3T3 mouse fibroblasts
- Chicken chondrocytes
- Pituitary cell cells
- Ovarian cell cells
- Q5B and Y8 transgenic mice
- Cortical neurons
- DRG (dorsal root ganglia)
- Neocortical cells
- Spinal cord neurons
- Hippocampal neurons
- Cerebellar granular cells
- GH3 pituitary tumor cells
- SF 21 insect cells
- GT1-7 mouse cells
Corning Poly-D-Lysine (PDL) microtiter plates are coated with PDL (molecular weight range 70 to 150 kDa). PDL provides a positive charge to the surface for better cell attachment.
- improves the differentiation of primary neurons, glial cells, neuroblastomas
- improves the adhesion of transfected cell lines, including HEK-293
- helps cells stay adherent during the test
Corning offers this surface for 96- and 384-well microtiter plates.
Corning Ultra-Low Attachment – polystyrene with a surface with low adhesion
This surface is suitable for:
- maintaining the cells in a suspension state
- study of tissue-specific functions of some cancer cells (ie MCF7 breast cancer cells)
- prevention of adhesion initiated by stem cell differentiation
- prevention of adhesion and activation of macrophages and neutrophils
- selective culture of tumor or viral transformed cells as unattached colonies (replacement of soft agar assays)
- growing primary cultures of tumor or adult stem cells as unprepared spheroids
- prevents anchorage-dependent cells, such as fibroblasts, from adhering and dividing
- No special procedures are required when using this surface
- reduction of adhesion by up to 99.8%
The Ultra-Low Attachment surface is a covalently bonded hydrogel layer that is hydrophilic and neutrally charged. Because proteins and other biomolecules passively adsorb to polystyrene surfaces by either hydrophobic or ionic interactions, this hydrogel surface naturally inhibits non-specific immobilization by these forces, thereby inhibiting subsequent cell adhesion. This surface is very stable, non-cytotoxic, biologically inert and non-degradable.
Corning offers untreated polystyrene containers and microplates for growing cells in stationary suspension or in other applications where reduced cell binding is desired. These untreated containers are sterilized with low doses of gamma radiation, which slightly increases the wettability of the surface. Because some of the transformed cell lines (e.g., CHO-K1) and macrophages adhere to and grow on these hydrophobic surfaces, Corning also offers an Ultra-Low Attachment (low-adhesion polystyrene) surface.
Unmodified polystyrene surfaces are hydrophobic and only bind cells and biomolecules by passive hydrophobic interactions.
Corning® Synthemax® II-SC
Synthemax substrate allows for scalable, multistage expansion of pluripotent stem cells in serum-free media such as mTeSR®, after differentiation into a variety of cell types, including retinal pigment epithelial cells and cardiomyocytes, as well as proliferation of various progenitor cell types.
Corning Synthemax self-coating substrate is a unique, animal-free, synthetic peptide based on vitronectin, which contains the RGD motif and accompanying sequences. It is a powdered peptide copolymer that dissolves easily in water. It is used as a surface coating to promote cell adhesion for various stem cell lines.
- easily soluble in water – ready to use
- it can be coated on any format of culture plastic, giving end users additional flexibility
Corning Osteo Assay
A unique three-dimensional structure that mimics the surface of bones in vivo , for in vitro testing.
- cell growth and differentiation of osteoclast or osteoblast precursor cells
- direct reading of osteoclast and osteoblast cell function and differentiation tests such as TRAP (tartaric acid phosphate) staining, rupture resorption and bone node formation
- targeted drug screening for bone cell activity
- real-time imaging for high-content analysis
- quantitative determination of enzymatic activity by sampling cell culture supernatant
This Corning Osteo Assay inorganic surface is an inorganic coating of crystalline calcium phosphate on a polystyrene board designed to mimic the bone environment in vivo . The surface is designed to facilitate a variety of bone cell assays, including immunofluorescence staining, TRAP staining, and surface resorption assays for osteoclasts. The surface is available in a sterile pack multiwell plates, 1 x 8 Stripwell Corning TM and is stable at room temperature.
This surface also offers a consistent and defined alternative to preparing dentinal or bone incisions, which reduces variability in your test system and leads to a more consistent test reading.