In the past cultureware has mostly been focused on culturing standard cell lines that have historically been easy to culture. However, with the explosion of stem cell culture and the now common practice of culturing many different cell types, cultureware has needed to evolve as well. Over the years, improvements have been developed to increase the success of cultureware and to provide a more hospitable cell-growing environment, including innovations like gas-plasma treatment and biological extracellular matrices. Yet there is still opportunity to improve cultureware and address some of the most common challenges cell culture scientists face.
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Check out our guest blog on The Cell Culture Dish written by our own Graziella Mendonsa, Ph.D.
A Guest Blog by Graziella Mendonsa, Ph.D., Product Manager at MIDSCI
In the past cultureware has mostly been focused on culturing standard cell lines that have historically been easy to culture. However, with the explosion of stem cell culture and the now common practice of culturing many different cell types, cultureware has needed to evolve as well. Over the years, improvements have been developed to increase the success of cultureware and to provide a more hospitable cell-growing environment, including innovations like gas-plasma treatment and biological extracellular matrices. Yet there is still opportunity to improve cultureware and address some of the most common challenges cell culture scientists face. Techno Plastic Products (TPP) is a Premium Swiss manufactured line of cell cultureware that is designed by scientists for scientists. Virgin plastics of highest purity and quality are poured with perfection into uniquely engineered molds to deliver the perfect flask, dish or plate for adherent cells to attach to. Suspension cells also get an updated housing with a conical Bioreactor that significantly enhances distribution of gas exchange and nutrition. Quality control is the number one priority for TPP, which includes the testing of each and every piece manufactured at their site.
Maintaining Standard Temperatures
Cell culture in general is treated as a great in vitro model system to assay signal transduction, cellular cross talk and responses to externally applied gene activators or inhibitors. Since cells in culture are sensitive to environmental stimuli and react accordingly for adaptation, it is essential to maintain their necessary vitals such as temperature and CO2 stable and non-fluctuating. Almost every lab with multiple scientists performing cell culture usually stacks their flasks, dishes and plates to save on space in their incubator. However, a key question to ask is “Are the vessels in the middle of the stack the same temperature as that on the top or bottom of the stack?”. The answer is a resounding “Probably Not!.” Therefore TPP has engineered a raised growth surface on their flasks, dishes and plates to avoid insulation of the adjoining vessels and has designed the vessels to have a stable footing for enhanced security in stacking. Maintaining Optimal Nutrition, Gas Exchange and Gene Expression The foundation of cell culture at the time of seeding in a flask or dish, undoubtedly sets up for consequent cellular viability and propagation during passaging and for consistent downstream gene reporting assays. Therefore how cells are initially thawed from frozen and plated during the first 2-3 days has a dramatic impact on cellular behavior downstream. Therefore controlling the flatness of the surface upon which cells adhere to reigns important to obtain consistent cell monolayers. Cells that are growing in layers are not only deprived of nutrition and the optimal gas exchange, but are also now transcribing uneven gene expression. TPP manufacturers one of the flattest growth surfaces in the market.
Evaporation of Media
Evaporation of media in 96-well plates appears to be one of the most common complaints from cell culture scientists. This is due to the overheating of the outer wells and a consequent temperature gradient across the well plate and across the stack of well plates in the incubator. This uneven heating can also lead to condensation build up which are breeding grounds for bacteria. The temporary solution generically recommended is to fill the outer wells with media alone or PBS. However the wastage of unused wells for cell growth and experimentation amounts to almost 40%. TPP plates eliminate this concern with its raised growth surface for optimal stackability and consequent even heating from all sides. Ultimately, a 96-well plate is used for multiple replicates of control and experimental treatments and with TPP plates, one can achieve that goal because surrounding environmental conditions such as temperature are no longer a contributor to variation.
Eliminating Dead Space
Focusing in on the flasks, it is strikingly obvious that TPP flasks are shorter in length. That was purposely designed to encompass the entire bottom of the flask as the treated growth surface. Therefore, with no wasted “dead space” in flask, the cell to media ratio is now optimally maximized. All TPP flasks come with an angled neck; this small design tweak serves as the best method to eliminate media accumulation in the neck and thus the consequent high risk of contamination. TPP also has filtered caps for their flasks that are designed to be hydrophobic in nature, so that incase media does slosh against the cap when the flask is either laid at the microscope or put back in the incubator, the risk of contamination is minimized greatly. The angled neck also provides full access to the internal corners of the flask for a serological pipette or scraper during trypsinization or cell harvestation respectively. Summary TPP dishes have ingenuity spelled all over it. From the unique grip ring on the edge of the dish that prevents accidental dropping to the internal clock orientation for visual quadrants during microscopy, TPP continues to invent upgrades to the cell culture dish that take cell cultureware from standard to ergonomic. TPP cultureware, designed by scientists for scientist, provide you with unique and innovative upgrades that translate to more consistent seeding, handling, treatment and harvestation of cells. Peace of mind included.
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Researchers at KU Medical Center identify a new target for treating pain
June 14, 2013
By David Martin
A hormone that has been used to control blood pressure may hold the key to a new approach for treating pain, according to research at the University of Kansas Medical Center. The hormone, angiotensin II, constricts blood vessels and releases a substance that causes the body to retain salt and water. KU neuroscientist Peter Smith, Ph.D., and his colleagues have found that blocking angiotensin II receptors prevents the increase in sensitivity that normally accompanies inflammation. The mechanics of the angiotensin II system's ability to regulate pain pathways was not appreciated before, Smith says.
The study has been published online in The Journal of Pain.
The finding is based on a previous study of estrogen that Smith, professor of molecular and integrative physiology and director of the Institute for Neurological Discoveries at KU, published in 2008. "We know that the hormone estrogen makes women more prone to many painful conditions, and we found that estrogen actually caused some pain-sensing nerve cells to grow," he says, adding that many chronically painful conditions are accompanied by an increase in pain-sensing nerves.
In looking at what genes were affected by estrogen, Smith and his colleagues were surprised to learn that nerve cells started making more of a protein that allows cells to respond to angiotensin II. This led the researchers to speculate that the protein, angiotensin II receptor type 2, or AT2, might be important to pain.
Tests on rodents confirmed the relationship. In the experiment, rats' hind paws were injected with an irritant. Exposure to touch and heat revealed the paws' sensitivity.
Some of the rats received a chemical known to inhibit AT2. In these rats, angiotensin II did not cause pain-sensing nerves to sprout, and paw sensitivity was the same as in rats that did not receive the irritant. In short, the AT2 blocker uncoupled the link between inflammation and the nerve response.
"It appears that AT2 is critical for the sensations of pain, and that AT2 receptor inhibitors are about as effective as morphine - but without all of the dangerous side effects," says senior scientist Anuradha Chakrabarty, Ph.D., the paper's first author. Zhaohui Liao, M.D., also contributed to the study.
The study, Smith says, confirms that the proliferation of pain-sensing nerves associated with chronic pain syndromes is meaningful. "More sprouting means more pain," he says.
The study is the first to shed light on the mechanism of overgrowth of pain-sensing nerves, Smith says. Inhibiting AT2, he adds, "represents one of the few drug therapies that actually is known to be directed at the underlying biological mechanisms that may be responsible for some aspects of chronic pain."
Chronic pain has been estimated to cost the U.S. between $560 and $635 billion in health care costs and lost productivity. Nonsteroidal anti-inflammatory drugs may not work for all patients and have the potential to damage the tissue of the gastrointestinal tract, kidney and liver. Opiates pose the risk of addiction and overdose and are unsuitable for long-term treatment of chronic pain.
An Australian pharmaceutical company is currently testing an AT2 receptor blocker in a clinical trial with patients with postherpetic neuralgia, nerve pain caused by shingles.
The KU study was supported by the National Institutes of Health grant RO1HD049615.
Article found at http://ow.ly/mcdSL