Induced pluripotent stem (iPS) cells show great promise for treating intractable illnesses through applications in regenerative medicine and new drug development. At Kyoto University’s Center for iPS Cell Research and Application (CiRA), more than 400 personnel conduct research under the leadership of Director Professor Shinya Yamanaka. This rigorous work is made possible by the as-sistance of technical staff. We spoke with Megumi Narita, Yuka Kawahara, Ryoko Hirohata, and Ito Miyashita, four lab techs in CiRA’s Department of Life Science Frontiers.
A desire to contribute to socially relevant research
CiRA employs a great many female lab techs. However in Japan, the proportion of women in science is still low. What inspired you to pursue science-related careers?
Narita: In elementary school, I was shocked to read about the pomato (a hybrid plant developed by cell fusion that produces potatoes in the soil and tomatoes on the branches) in a science magazine. In college, I majored in molecular nutritional science in the Faculty of Agriculture. When I graduated, it was a difficult time for job searches, and I could not find a post in science. For a time I did quality control at a metal parts manufacturer, but I could not give up on my dream. When Professor Yamanaka moved to Kyoto University, new tech positions were announced, and I started working here in November 2005.
Kawahara: My mother greatly influenced my career choice. She used to purchase and read popular scientific monthlies such as the Newton magazine, which I was also fond of reading. In 2003, I heard the news that the human genome had been successfully mapped, and it made me very interested in genetics. I was hoping to do something that would serve the world of the future. I joined CiRA in April 2013.
Hirohata: Oh, I didn’t have a noble purpose like these two [chuckles]. I have been fascinated by dinosaurs since childhood. I entered university to study biology, thinking that it would be the fast track to dinosaur research, but later found that I also needed some liberal arts credits to become a museum curator [laughs]. In the film Jurassic Park they mined the amber to find viable dinosaur DNA to use for cloning, a scene that intrigued me. After finishing my master’s, I began working as a tech at Osaka University. While there I got married and gave birth to a baby.
Just when I was looking for a way to keep working and raising my child with the help of my parents, CiRA was established, and it advertised lab staff recruitment. I have been working here since August 2010. I was really lucky to find a job that suited my ambitions and circumstances.
Miyashita: In my case, I spontaneously developed a desire to become a researcher because my father and one of my aunts are corporate scientists. The Great Hanshin-Awaji Earthquake occurred when I was a high schooler. I witnessed food completely disappearing from shops. With Japan’s low food self-sufficiency, one full-scale disaster would place this country in complete chaos, I figured. As I studied agricultural science at college, I realized that disease protection of farm products would be more effective in increasing harvest than time-consuming improvement through breeding. Consequently, I majored in plant pathology in my graduate program.
Plant pathology is different from the area where you currently work, is that right?
Miyashita: When I completed my graduate course, there were few promising job opportunities. For a while, I had temporary tech staff jobs in corporate research departments. I started working here in October 2010. As Ms. Narita was about to take maternity leave, I was asked to replace her. I decided to accept the offer because I could leverage my knowledge and expertise, including molecular biology techniques.
Precision and persistence required
What does your routine tech work involve?
Narita: Ms. Kawahara, Ms. Miyashita, and I belong to the research group headed by Professor Yamanaka, the biggest lab at CiRA. I am with the subgroup working on iPS cell–derived myocardial and blood cells intended for use in transplantation. For example, I prepare plasmids (DNA fragments to be transferred into the extranuclear cytoplasmic compartment of the iPS cells that proliferate independent of the chromosome cycle) to produce myocardial or blood cells. Checking the genetic profile of differentiated cells is also my responsibility.
Kawahara: I work for the group that investigates the mechanisms underlying iPS cell formation. I am mainly involved in the chemical screening process. Its objective is to discover alternatives to specific components that will help improve the responses of a variety of cells and thereby reduce the cost of culture media. We screen thousands or tens of thousands of compounds robotically or manually.
Hirohata: You have to just do whatever comes your way, right?
Kawahara: Yes. We test an enormous number of compounds, but oftentimes we come up with no positive results no matter how many runs we may make. Sometimes, several weeks pass without any significant results, but other times, we get a sequence of positives.
Narita: I find it is sort of like betting.
Kawahara: My basic attitude is that it won’t work out as easily as I wish. That’s what makes me very happy when I discover a useful compound.
Will your work lead to future clinical trials?
Miyashita: I am assigned to the project team to stock iPS cells for future regenerative medicine use. I participate in quality checks to evaluate the stock cells produced in the cell preparation facility. I need to be very careful. Since my section is located downstream of the entire testing process, my schedule becomes tighter if upstream tests are delayed. There is not much I can do to prevent that, and I feel pressure to complete my part as planned because the deadlines cannot be altered.
Hirohata: I am assisting Associate Professor Knut Woltjen, whose research is focused on the use of genetic information for generative medicine. Dr. Woltjen is good at building Lego models. He designs the precise details of plasmids like Lego models, saying, “Let’s have this gene illuminate green, and let’s generate that protein.” My responsibility is to create plasmids according to Dr. Woltjen’s designs. I also conduct experiments to investigate how the plasmid-derived iPS cells differentiate in mouse models. I occasionally receive requests from members of separate groups, asking me to create certain plasmids. When I learn that an experiment was successful, or when I see pictures of the cells transfected with the plasmids I made, I am proud of what I did, knowing that my work served a pur-pose.
It seems that you engage in a lot of precise and sensitive operations. Do you think you are cut out for such work?
Narita: I do enjoy it.
Kawahara: I am very into it, too.
Hirohata: I am a laid-back type of person at home. My family seems to understand and accept that I need to be so at home because I must be so precise at work.
My impression is that patience and persistence are necessary traits for techs. What do you think?
Kawahara: I think I am very patient and persevering. When the results are not favorable, I keep hanging on until I obtain positive results.
Narita: When my work is not going well, I believe that there is a cause for that. I like to try out this or that option, considering a variety of available remedial measures.
Hirohata: Toughness is important; so are communication skills. Because our job is to assist the principal investigators and other senior scientists, we must not assume a “know-it-all” attitude. I communicate as frequently as possible with my boss to understand what he wants from me.
Business as usual the day after the Nobel Prize was announced
Professor Yamanaka received the 2012 Nobel Prize in Physiology or Medicine together with Professor John Gurdon. All of you were at CiRA at that time. What were your reactions to the news?
Narita: Although iPS cells were not yet in the stage of making tangible social contributions at that time, Dr. Yamanaka was awarded the prize. I was surprised, purely happy, and felt great.
Hirohata: The prize was given only six years after Prof. Yamanaka first created the cells, and such a short time interval represented the weight of expectations for iPS cells, I thought.
Miyashita: My parents, relatives, and friends told me that it was a fantastic event. Knowing that CiRA captured a tremendous amount of attention around the world, I also felt pressure and tension as one of the employees.
Were people at CiRA excited?
Miyashita: No, the atmosphere was surprisingly calm and business-like.
Narita: The announcement was made on a Sunday, and most people learned about it through spot TV news coverage. We saw that, in other countries, lab members often gather together and celebrate the prize winner with wine. On my way to work the next day, I wondered if we would hold a party like that. However, it was just another ordinary day.
Miyashita: There was hardly any merry-making.
Narita: I personally wanted it to be merrier [laughs], because I knew the cozy, family-like atmosphere of CiRA before it became the big organization that it is today.
Miyashita: Above all, Dr. Yamanaka is a very modest and low-profile person, and his down-to-earth attitude encourages us to work incessantly to seek clinical applications of iPS cell technologies.
Escherichia coli, lovable lab partners
Can you tell me about some of the special skills or habits that you have developed as techs?
Hirohata: When I take sterilized tubes out of a container, I must do so without directly touching them by the hand. By adjusting the gradient of the container and the level of physical force applied, I can pop out the exact number of tubes I need.
Narita: That is a wonderful skill. I remember that I was measuring 500 mg worth of a chemical agent. When I poured the powder out of the container, it was just the amount I needed.
Hirohata: Everyday practice help us develop a full command of certain skills that enable us to be sensitive to abnormalities. For example, when using a micropipette to fractionate liquid, I sometimes notice that something is wrong with the fractionation, feeling that the 1 mL aliquots just prepared are somewhat different from those I commonly do. When I send the device in for maintenance, they often report malfunctioning of a spring or other parts.
Miyashita: I make it a habit to completely clean the cell culture workbench to prevent contamination by dust or microbes. Consequently, I am not satisfied at home unless I completely clean the kitchen table before eating. When the chopsticks I use for cooking touch things unrelated to cooking, I automatically think, “Oh, I am risking contamination” [laughs].
Hirohata: I often chat about Escherichia coli with my colleagues. Those microbes are indispensable research partners, although people in general consider them bad and ugly. When preparing plasmids, we transfer them into E. coli for multiplication. They are very helpful because they can proliferate rapidly.
Narita: When I hear people speak badly about E. coli, I am tempted to tell them that these microbes are good and that we owe a lot to them [laughs]. The E. coli strains we use have been developed for specific research purposes. I have a passion for them.
Hirohata: So do I.
A Mission to Meet the World’s Expectations
Fast-paced research efforts are being made toward clinical applications of iPS cell technologies. What are your thoughts and future hopes?
Miyashita: I am currently involved in a project to stock iPS cells for future regenerative medicine use. I feel an enormous responsibility. It will take some more time before these technologies become clinically applicable. I am determined to do my current work correctly and precisely to see the day when iPS cells are used to cure diseases.
Hirohata: CiRA receives donations from a wide variety of supporters including patients with refractory diseases. I sometimes have the occasion to meet such patients in person, when they visit us. This renews my commitment to our work. I wish to contribute to yielding positive results by giving it all that we have.
Narita: I wish to help society by contributing to clinical applications of iPS cell technologies. Although I sometimes wonder if I have what it takes to meet expectations, I am ready to do my best to help us make a breakthrough.
Kawahara: I am giving my current work my very best. I wish to improve my technical skills so that I can help patients.