South Dakota State University has long had a reputation for involving students in crop research. But what is being grown in a small room on the top floor of the pharmacy building at State goes beyond traditional row crops or even exotic plants.
Three undergraduate students are working with two assistant professors in the College of Pharmacy to grow human brain cells. It is one of a handful of places in the world where the work is being done, according to the husband-wife research team of Suman Mukherjee and Manisha Sonee.
Although both work with and help to grow the cells, their research falls into separate areas.
Mukherjee experiments with a compound that could be used in the prevention and treatment of Alzheimer's and Parkinson's diseases, which affect the brain. Sonee seeks to better understand the mechanical functions of cells when they are exposed to Alzheimer's Disease conditions.
Stacy Peters of Pipestone, Minn., says each of the students work on both projects.
Peters, who started work late last summer, says she was intrigued by the topic. "The fact that they were human brain cells and the work is concentrated on treating and preventing neurological diseases caught my attention."
Fellow students Carisa Finke of Pipestone and Stephanie Villmow of Delmont, S.D., got a head start on Peters in their work. Finke started in January 2001; Villnow started last May.
As a result of a proposal that each student wrote on their role in the project, they were selected for scholarships that entitled them to work with the researchers.
"I love it. It's fun," Finke says of the project. "I really wanted to get this project because I've always been interested in Alzheimer's Disease and that's what my project primarily involves."
Villmow reports that "Carisa had started working already" with Mukherjee and Sonee. "She was telling me the neat things she was doing, and I didn't have plans for the summer. I thought I should do research. It's probably a once-in-a-lifetime possibility."
Mukherjee says the students, who are in their fourth year of the six-year PharmD program, are an integral part of the project. "We have to spend quite a bit of time with them to do the techniques correctly," he says, "but once we do, we rely on them to do it all."
He describes the students as "first rate. They don't look at it like a job. They look at it like a challenge, and that takes it to a higher level. I have been amazed by their grasping of these different concepts, how they do the experiments, their hard work, and general enthusiasm.
"None of my students have had prior research experience. They start out in the summer with varying degrees of apprehension, but by the end of the summer, they're usually working Saturdays and Sundays."
The research is not required for the students to receive their degrees. But Finke and Peters are both considering careers in pharmaceutical research and know their work with Mukherjee and Sonee could help them solidify their post-graduation plans.
Finke first experienced undergraduate research in summer 2000, when she collected data on cheese and milk for David Henning, an associate professor in dairy science.
Villmow says that learning the techniques needed to grow brain cells isn't easy. "There is a lot of practicing your technique with water." After about a month, she was prepared to feed the brain cells. But after ten weeks on the job, she wasn't prepared to do the volatile task of dividing the cell cultures.
It took about six months for Mukherjee and Sonee to "get a handle on" how to grow human cortical neurons, the cells found in the front part of the brain where a portion of the memory function occurs, Mukherjee said.
"Human brain cells are very difficult to grow. By nature, they're resistant to duplicating. We have to feed them and coax them into dividing," says Mukherjee, who started working with the cells shortly after he joined the SDSU faculty in fall 1999.
"Brain tumor cells are easier to grow, but Alzheimer patients most likely do not have brain tumors," he says.
The researchers, who also have a teaching load, find their schedules "completely dictated by the cells," says Mukherjee, who, like his wife, holds a bachelor's degree from Jadavpur University in Calcutta, India, and a doctorate from the University of Southern California.
Sonee notes that the cells "need to be fed every day, and when you're working with them, you have to be very gentle with them. You have to maintain them in certain conditions" -- just the right temperature, humidity, and carbon dioxide level.
"And they have to be at the right concentration. There can't be too many or too little," she adds.
The cells are fed a commercially-made liquid nutrient mix. Although an experienced scientist can feed the cells in an hour, providing a serving of "brain food" to the cultures is a bit more complicated than squirting an eyedropper of the red fluid into a fish tank.
Sterile conditions are the key.
The room, formerly a prep area for a student lab, is equipped with $61,000 worth of instruments for cellular research. A key component is the biological safety cabinet, which has an ultraviolet light to kill microbes that would feast on the cells.
To further eliminate microbes, the cabinet and all instruments are wiped down with a seventy percent ethanol solution.
Operators must wear gloves and be careful not to let their hands pass over the small plastic flasks where the cells are being grown. Until the researchers are ready to start growing the cells, the vials of commercially purchased cells are kept in liquid nitrogen.
After research on a particular batch of cells is completed, the sample is refrigerated at --eighty degrees to keep enzymes from making further changes to the cells. The preservation technique allows for later research on cells, Mukherjee said.
Before the cells are fed, the nutrient is heated to thirty-seven degrees centigrade, which is body temperature, he said.
The work is exacting. "We just can't afford to have these cells die on us because if these cells die, you can easily lose a couple months of work in two minutes," Mukherjee said. So far, the couple has had only one major cell die-off, he said.
Growing the cells is a science in itself, but that only starts the work of Mukherjee and Sonee.
He uses the cells to better understand why cells die, as they do in Alzheimer's and Parkinson's diseases, and what can be done to combat that. Mukherjee has found that a compound using nicotinamide, a vitamin which he purchases, is effective in preventing cell death.
Nicotinamide converts to ATP (adenosine triphosphate), which provides the cell its energy, he said.
"Our brain is separated from the rest of the body by a barrier. It prevents poisons from getting into the brain. Most of the drugs we take cannot get into the brain, but nicotinamide is a compound that can get into the brain," Mukherjee explains.
Sonee's work examines both healthy brain cells and those damaged by conditions that mimic Alzheimer's Disease.
By gaining a greater understanding of how motor proteins work and how the transportation system within a cell functions, she hopes to give researchers a platform from which they can better seek a cure or find a cause for Alzheimer's Disease, she said.
Eventually, they hope to gain funding from the National Institute of Health or other national organizations for their work.
In the meantime, the College of Pharmacy's research "farmers" diligently grow the cells that are needed to help solve two of the world's most tragic diseases.
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