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Perspective On: A Cell Culture Lab

Many studies suggest that toxins in the environment may be a cause of autism, affecting a mother’s unborn child as it develops. However, proving this for certain is difficult, as it is something that cannot be tested directly.

Rachel Muenz

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Discovering the Secrets of Brain Development - In a Dish

Passion and persistence push this lab forward as it searches for the cause of autism

“You can’t feed a toxin to a pregnant woman and say to her, ‘Hey, we just want to give you these toxins and see what happens to your child.’ Obviously that can never happen,” says Dr. Chris Fasano, a principal investigator (PI) at the Neural Stem Cell Institute (NSCI) in Rensselaer, New York. “But with stem cell technology, we can model [that] in a dish.”

Fasano’s lab at the NSCI uses stem cells to gain a better understanding of how the human brain develops, even in the earliest stages, aiming to discover what causes autism. “Once we understand that process, we can try to understand what happens when things go wrong,” he says.

As a whole, the NSCI is focused on using stem cells to understand and create new therapies for nervous system diseases, including blinding disorders such as macular degeneration and neurodegenerative diseases such as Parkinson’s and Alzheimer’s.

Fasano’s roughly 1,000-square-foot lab is staffed by five employees, most of whom are students working toward their master’s degrees or doctorates.

“When the majority of your lab [staff are] students, it’s important as a manager or principal investigator … to remember that the primary [goals] of that relationship [are] education and training,” Fasano says. He adds that stem cell science is a very competitive field and students tend to want to move quickly and publish as many papers as possible. As a primary investigator, he says, while it would look good for the lab to have students publish often, it’s more important that they are properly trained in basic science. “I think that’s a challenge nowadays in a very competitive environment where you want to push your students to produce but you have to be cautious because they are students and they are learning and you have to make sure that they are properly trained first and foremost,” he says.

In his lab, that training starts with basic cell culture techniques, including aseptic technique and how to grow and maintain cells. Students also learn a bit of molecular biology, dealing with DNA, cloning, polymerase chain reactions (PCRs), immunochemistry, and staining cells with different antibodies.

“They all receive a similar level of training,” says Fasano, who has been at the NSCI for about four years and is also director of research and development at the institute. “Some students might, for their project, have a specific application that they have to learn … but everybody gets a fairly thorough cell training course and basic molecular analysis skills like running a DNA gel and doing a PCR.”

Those skills help staff deal with three to four main projects at any one time, along with a number of side projects. “All are part of a common goal,” Fasano explains. “There might be four projects that are different, but they’re all centered upon trying to understand something in the realm of human brain development.”

Balancing his staff ’s individual needs while also promoting teamwork is another management challenge Fasano faces in his lab. “It’s like I have a bunch of kids,” he says. “You want to empower them with a sense of individuality; you want to give them their own project that they feel is theirs, but at the same time you want them to collaborate and work with their labmates because they are there for a common goal.”

The manager’s chair

Fasano says one of the most difficult parts of transitioning to a lab management position is learning how to manage people successfully.

“When you receive a PhD, in your doctoral training nobody trains you on how to manage anyone. It’s not part of grad school,” he says. “There’s no class on management, no one’s coming in there to teach you how to run a business. It’s purely science.”

Those skills come through experience, but having a mentor is something that Fasano says helped him a lot when he became a manager.

“Having a mentor in someone who’s been managing a lab or has been in a company for a while to talk to and ask their advice on how they did or didn’t do it successfully really helps,” he says. “I’ve been lucky to have mentors for my PhD and for my postdoc who’ve been great in giving me advice for how to successfully manage.”

He says the most important ways he motivates his staff are through creating a positive work environment and choosing the right people who are willing to work hard as a team.

“You have to bring people into your lab [who] are selfmotivated and really want to push things forward,” Fasano explains. “You can teach anyone how to do an experiment, but you can’t teach someone how to be motivated.”

He adds that he works to create a good environment for staff by being passionate and interested in the work they are doing.

“In science, 90 percent of the time experiments don’t work,” he says. “Ninety percent of the time your grants don’t get funded. So as a manager, my job is to always be positive, to always keep the morale up by trying to inspire them and say, ‘Your work matters. Your data really is going to make a difference.’ That creates an environment where people are much more upbeat.”

As PI, Fasano is in the lab about two to five times each day. In an average day, staff will feed their cells, analyze them, chat with Fasano about their latest data, and analyze that data.

“I like to sit down and have a dialogue with all my students and techs,” he says. “I like to get them to think, get them to ask questions. If they have an idea, I want them to tell me what it is and I want them to tell me why they think it’s a valid idea.”

Apart from the management challenges, Fasano’s lab faces many of the same issues other labs are facing— running a lab on reduced funding and disappearing grants. He says passion and persistence are the keys to overcoming such negatives.

“If you’re going to be a manager in this lab world, whether you’re going into industry or academia, you have to have a tough skin and a tough stomach,” he says. “Your passion for it has to be so strong that in the face of many critics, you’re going to keep going. A lot of people say that science is just not viable as a long-term career anymore. I disagree with that. I think it just takes the right person. You have to keep going and keep trucking through, and eventually your ideas will pan out.”

For Fasano, the positives of the job are also similar to those of many other managers—having the opportunity to learn and do something new each day, doing challenging work, and working with young scientists.

“My role in this as a manager and as an investigator is not just to teach them science but also to be a mentor to them in their career and their life at this stage,” he says. “I need to establish relationships with them and get to know them as people and help guide them through this stage of their life and onto the next. That involves a lot of responsibility, and it’s great to work with really smart, young people.”

Getting technical

On the technology side of things, Fasano says reagents are especially important in stem cell culture work, so much so that NSCI created its own reagent company called StemCulture so that it would have full oversight of the quality control process.

“When we grow cells, especially stem cells, they require a lot of care,” he explains. “They’re very sensitive, and they require a daily cell media feed, so you have to come in every day and feed them. That’s really annoying because it’s very tedious.” He adds that if you get a bad reagent in cell culture, that can destroy a lab’s cells for months, taking down its whole process as well as team morale.

To solve this issue, StemCulture created a growth factor product called StemBeads, essentially a time-release stem cell food.

“You just add it to your dish and it will feed your cells for you, so we don’t have to constantly feed cells every day,” says Fasano, who’s also chief operating officer of StemCulture. “In a way, we’ve created a management tool for cells. We’ve created a way to manage cells less intensively. It allows the students and the postdocs to focus on more important things.”

His lab also uses a lot of high-throughput RNA sequencing technology in order to better understand human brain development from start to finish. That technology “looks at all the genes in our genome at static points, and then we can decode that to get an idea of what genes go up and what genes go down as the brain develops,” Fasano says. “Then we compare that to samples from autistic patients to see how those gene changes are now different.”

Induced pluripotent stem cell technology (iPS) plays a strong role in the work of Fasano’s lab as well, allowing researchers to “take a sample of somebody’s skin and reconvert that all the way back to an embryonic stem cell,” he explains. “This allows us to generate stem cell lines from autistic patients that we can then turn into brain cells and see how that went wrong.” The combination of those technologies gives the lab a powerful model to understand how the human genome changes in people with autism.

However, the two technologies are not without their difficulties. With iPS, there is a lot of variation each time the lab makes a cell line, so they have to use many experimental controls to account for that variability, while with RNA sequencing the challenge is making sense of the huge amount of data gathered.

“You really need the help of computer scientists and mathematicians to help you decode that to have it make sense,” Fasano says of that data. “As biologists, we’re not trained to do that, so you have to be interdisciplinary. You have to work with people outside your field to make sense of the data you’re getting.”

Overall, new technologies, including cell culture kits, are making it easier to grow cells better and more efficiently, he says. Admitting he may be biased, he says the StemBeads technology has really helped, as researchers have gone from needing to feed cells every day, even on weekends, to two or three times a week.

Main Technologies Used in the Lab

  • StemBead growth factors
  • High-throughput RNA sequencing technology
  • Induced pluripotent stem cell technology

“It’s important to be in the lab, but it’s important to be out of the lab,” he says. “You want to be dedicated, but you need some time to yourself. This kind of advance has allowed that to happen without sacrificing quality of cells.”

He says StemCulture aims to grow the StemBeads product line, and he also hopes to grow his own lab, get more funding, and add some postdoctoral fellows to his staff to offer extra guidance and support for the grad students. All that will help them continue their promising work.

“The fact that we can now take an everyday-occurring toxin and ask basic questions like ‘What does it do to early brain development? How does it affect the genes that are changing during what would be the equivalent to pregnancy [in a culture dish]?’—it’s really exciting, and we think there’s going to be a lot of cool data there that’s going to elucidate [autism] and have a really positive effect on the general health of people going forward,” Fasano says.