Making the ‘best’ better

Media system extends lifespan of functional neurons to advance neurodegenerative research

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WALTHAM, Mass.—Scientific and medical researchers are making progress in understanding brain function, how neurons transmit and receive information and how they factor into the presence and/or susceptibility to neurological diseases such as Alzheimer’s disease, multiple sclerosis and Parkinson’s disease. Since 1993, Thermo Fisher’s Gibco B-27 Supplement and Neurobasal Medium has been the standard for a variety of neuronal culture applications, with citations in more than 11,000 publications. The customer base has included researchers studying neural physiology, toxicity and therapy, as well as those involved in the discovery and use of pluripotent stem cells.
The inability to keep neurons alive and functioning as they do in the brain for extended periods of time has been a serious challenge to fully understanding how the brain functions. For example, rodent neurons typically only survive in current media for around 14 days, and their health degenerates significantly during that period. Functional in-vitro studies of neurons and neural networks require that the cells are maintained for prolonged periods of time at an optimal density.
For primary rodent neurons, full functionality and maturation requires maintenance in vitro for approximately 21 days. As the desire for more reliable and biologically relevant models has increased, so too has the necessity for a next-generation media system that can maintain and mature optimal densities of functional neurons over longer periods of time in vitro.
The result has been a concerted effort to improve cell culture media to enable neuroscience researchers to experiment for longer periods of time with neurons in a dish that look and function like neurons in the brain—without neuron degeneration. Such an advance will allow them to study processes that were not possible before, including the therapeutic potential for different drugs in neurodegenerative diseases.
While the core formulation has not undergone too many substantive changes, Thermo Fisher “thought there was room for improvement in maintaining the health of cultures outside of the intact brain, to create in-vivo-like physiology in a dish,” according to David Kuninger, senior research and development manager. Laura Requena, director of product management, primary and stem cell systems, added, “We wanted to make the best better, to maintain healthy, long-term cultures.”
To meet this need, the new Gibco B-27 Plus Neuronal Culture System was developed to enable superior modeling of neurons, fundamental to advancing scientific understanding of the brain and neurological disease. The B-27 Plus Neuronal Culture System is a next-generation media system that provides the highest rate of in-vitro survival of primary rodent and human stem cell-derived neurons. It features an optimized formulation, upgraded manufacturing process and more stringent quality control for raw materials and final product.
According to the company, the B-27 Plus Neuronal Culture System enables neuroscience researchers to experiment in a dish with neurons that look like and function like neurons in the brain. The system allows researchers to study processes that were not possible before, including the therapeutic potential for different drugs in neurodegenerative diseases like Alzheimer’s.
“For drug discovery and development, the B-27 Plus, in allowing for better survival and activity of neurons, gives scientists a chance to use fewer neurons and get higher-quality data with cell health maintained longer, more robust activity, improved markers of maturation and the ability to create disease-specific models,” Kuninger said.
Requena added, “For the neurophysiologist, there is the ability to do more with current cultures. They can survive longer and have more robust activity. Tissue is precious, and we can do more with less cells. We can isolate cells and be sure they survive.”
Improvements in the B-27 Plus system include increased neuronal survival by more than 50 percent, resulting in a more competitive cost for the same number of neurons compared to other products. The B-27 Plus Neuronal Culture System promotes the highest survival rates of primary and stem cell-derived neurons in both short- and long-term culture, which enables successful downstream applications using these cells.
Another benefit is accelerated neurite outgrowth in short-term neuronal cultures. The B-27 Plus Neuronal Culture System improves the electrical activity in long-term cultures relative to the classic B-27 supplement and neurobasal medium. The accelerated neurite outgrowth and increase in spontaneous firing, together with a concomitant increase in synapsin staining for neurons grown using the B-27 Plus system, indicates the formation of functional synapses. Finally, the culture system is said to seamlessly replace other neuronal cell culture systems, such as classic regular versions of B-27 Supplement and Neurobasal Medium, with no changes to current workflows.
“The B-27 Plus Neuronal Culture System is a robust system that provides improved cell viability, high-quality electrophysiological activity, interconnectedness of cultures and reproducible data,” Kuninger summarized.

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