The ghost of cells past: Part 1

 

Editor’s note: We are pleased to present the first episode of a new four-part story by Deborah Flusberg, about a lab research project that suddenly gets personal.

When people asked Robert why he wanted to go into biology research, he never mentioned his sister. He didn’t talk about the week when she had gotten a high fever and had to be taken to the hospital, staying there for what felt like months; the headaches and the blackouts. Nor did he bring up what had happened later, when she’d had to stay home from school and her hair had fallen out. At first, it had seemed like everything would be okay, that she would be cured. Robert didn’t like to think beyond that point. It was all a blur, like a distant film reel playing over and over in his twelve-year-old memory – one that he would rather forget.

Robert had heard about things like this in other labs: competition so great that people sabotaged each other’s experiments

Even when close friends from his childhood brought it up gently, noting that his chosen career path was such a wonderful way to honor her memory, he just looked at them and said nothing. In his mind, the two were not connected. Robert liked asking questions, trying to understand how things worked. This was what had driven him to major in biology, to apply to graduate school, and to aspire to be a researcher – to seek an understanding of life’s great mysteries. Not to honor anyone’s memory. And not to try to help sick children.

He really didn’t want to study cancer – in fact, he had tried to stay as far away from it as he could. This had worked well enough in the small undergraduate college that he had attended, where basic science was praised above all. But once he got to graduate school, he found that it was everywhere, this web of cancer, casting its net upon even the most reluctant of researchers. When he had told one of the professors that he’d interviewed with that he wasn’t interested in studying cancer, the professor had laughed.

“If you want to get funded,” he had said, “you’ll have to study cancer.”

Anything that had to do with the growth of cells, their structure, life-cycle, even their development within a tissue, was all related to cancer, because cancer represented the case when these processes started to go wrong. And in order to get research money, said the professor, you had to study everything that could possibly go wrong.

But he could study fruit flies, Robert had responded. Researchers who used the fruit fly Drosophila melanogaster as a model organism usually focused more on questions of organismal development, rather than cancer. For example, he knew from his undergraduate days that researchers would breed flies with different genetic mutations and characterize the effect of the mutation on the body shape or behavior of the fruit fly – the mutation’s ‘phenotype’. Or maybe he could study neurodegenerative diseases – there was money in that, and it wasn’t cancer.

Robert’s professor had just smiled politely when Robert suggested this, not saying yes or no. “You are an idealistic young man,” the professor had responded. “And that is always a good thing.” And so Robert’s disdain for cancer had helped him to get into graduate school.

In the first year of the program, the students in Robert’s class had to rotate through three labs before choosing the one they would ultimately join for their graduate thesis project. There was a thick book of labs to choose from, but his professor had been right: most of them were linked to cancer. Robert was able to find two labs that piqued his interest that were not related to cancer research, and so he did his first two rotations in a Drosophila lab and an Alzheimer’s lab. But when it came time to choose his third rotation, toward the end of his first year of graduate school, Robert didn’t know what to pick. The Drosophila lab was a very small group, and the Alzheimer’s lab was large and impersonal. Robert wanted to find something in between. There was a professor whose class he had taken the first semester who was a dynamic speaker and with whom Robert thought it would be exciting to work. The lab was studying something called “cellular stress” and used all kinds of imaging and biochemistry techniques to address these questions. But their questions were directly tied to cancer.

Robert decided to rotate there anyway. He didn’t want to be too picky and the lab seemed like a good one, had gotten positive reviews from his other classmates. It was a relatively big lab with 15 or more members, but that meant that no one minded having an extra person passing through, and Robert was assured that everyone there was happy. And it sounded like he could pretty much pick his own rotation project, as long as he studied something related to the lab’s general area of research.

**********

Robert’s days in the lab were long. He would get in late morning and set up an experiment before going to eat lunch. The bulk of his experiments, though, took place from the afternoon until late at night. It was how he liked it – the lab was quieter at that time. He could think, without too many outside distractions invading his brain.

During his first days in the lab, Robert learned how to take care of cells: how to thaw them from the lab’s stock of frozen vials into a petri dish filled with growth media; how to prepare such media, one 500 ml bottle of nutrients plus 50ml of a tube of bovine serum to aid cell growth, plus a small amount of antibiotics to prevent bacterial contamination of the cells. He learned how to “split” or “passage” the cells when they started to grow crowded in the dish: to break off their attachment to the dish using an enzyme called trypsin, and then once the cells detached, to collect them in fresh media and add a small fraction of them to a new dish. Then to let those cells grow until the dish, once again, became crowded. The fact that these were cancer cells didn’t bother Robert too much: he just decided not to think about it. In his mind, they were human cells, in all of their wonder and glory. There was, he knew, so much within them to be discovered. And it was easier to do these studies in cancer cells, he had learned, because of their robustness and rapid growth compared to “normal” cells.

In terms of the stress response that Robert’s new lab was studying, the usual cells that the lab worked with were giving uninterpretable results, ones that were different from those that another lab in Japan had published. Kelly, the postdoc that Robert was working with, decided that it might be better to try another cell type, preferably one that was more representative of glioblastoma, the lab’s new interest. The old cell line was one that many labs the world over had been experimenting on for decades, and during this time it had mutated so much that it was not really representative of any particular cancer anymore. Moreover, stocks of this cell line likely differed from one lab to the next, due to its high mutation rate and abnormal number of chromosomes, which tended to rearrange themselves in all possible manner of combinations during cell division. The lab down the hall had been studying glioblastoma for some time and had various newer cell lines in their liquid nitrogen freezer stock. One of the postdocs there recommended a particular cell line that he had worked with the year before that was fairly easy to grow. No one in the lab, as far as he knew, was working with the cell line now, so Robert was free to take as many vials as he liked.

The cells were called GL-350. They were star-shaped, with a partial fibroblastic morphology, and the postdoc had been right, they grew pretty well in the dish. The media was a bit complicated to make up, with a bunch of supplemental factors not required for the other cell line he had learned to work with, but the postdoc was kind enough to provide Robert with his stock of aliquots.

Robert got into a routine. He would plate cells in the evening to perform his experiment on them the next day. The experiment usually involved treating the cells in some way to induce or prevent a stress response, and then collecting the cells to measure what was going on inside them, like changes in the levels of certain proteins. Robert liked the work. When he ran a western blot, a technique that allowed him to detect the levels of specific proteins within the cells, he imagined himself to be an explorer, only instead of seeking land and adventure, he was seeking knowledge and understanding. “Proteins of the Wild West” was the title of the movie that he starred in within his mind. He pictured himself with a cowboy hat, saving science from imminent doom. Riding a horse through the desert, with cartoon proteins as his companions. In the Drosophila lab, he had picked up names of genes that he felt a connection to: Wingless, Hunchback, Son of Sevenless. Now, the proteins accompanying him on his journey had more obscure names: p38, Hsp70, MAP Kinase. But these “aliases” only made them seem more interesting to Robert, who was determined to uncover their true identities.

Everything was going well. And then, just as suddenly, it wasn’t. Robert came into lab one morning feeling excited about an experiment that he was planning to run. When he arrived in lab, he went straight to the incubator where he kept his cells growing. Only when he opened the incubator door, he did a double-take – his cells were gone. At first he thought that he was mistaken, that maybe he had put them on a different shelf in one of the other incubators. But he looked in all of the other incubators, and his cells weren’t there either. Robert asked everyone in the lab whether they had seen his cells, and no one had. He sent an email to the department, asked the postdoc in the other lab who had given him the reagents. Nothing. He wondered if he was going crazy. He was sure he had been there the night before and plated cells. But what if he hadn’t? Had he dreamed it? Or slept-walked back to lab and did something with his cells?

Or what if someone in the lab wasn’t telling the truth? There was no one that he thought of as particularly “suspicious”. Could one of the cleaning staff have thrown away his plates accidentally? Robert searched all of the biohazard trash cans, which were still full that morning, but could find nothing. Was someone in the lab lying? But why would anyone want to steal his experiment?

Robert had heard about things like this in other labs: competition so great that people sabotaged each other’s experiments. But he was just a rotation student, with barely any results to show for himself yet. He certainly wasn’t a threat to anyone. And the cells he was growing were nothing special – anyone could get a plate for themselves if they wanted to – all they had to do was ask. In fact, no one within his own lab was studying these particular cells at the moment – so it seemed unlikely that they would have taken Robert’s plates by mistake. They had been clearly labeled with his initials. Robert had no idea what to do.

He decided to just go ahead and thaw a new batch of cells, and within a few days he was able to set up his experiment again. He made up a fresh bottle of media, and spent the evening plating. As the week went by, he convinced himself that he must have been mistaken about the cells from the previous week. He had been working so hard, staying late in the lab; maybe he had fallen asleep and only dreamed about plating the cells. Or, perhaps one of the cleaning staff had accidentally knocked over his plates and thrown them out somewhere else, and was too embarrassed to come forward. In any case, it was not that much work to set it up again.

The next morning Robert came in and checked the incubator. Thank goodness, cells in place, growing fine. Everything had been seeded at exactly the optimal density, and Robert was pleased with his progress in technique and skill. He went to the tissue culture fridge to start taking out the reagents he would need for the day’s experiment. The first thing he reached for was the fresh one-liter bottle of media that he had prepared the night before. He had only ended up using a little bit for plating his cells, perhaps 100ml, and the remaining 900ml was left to use for today’s experiment and hopefully, for the rest of the week. But when Robert reached his hand into the fridge, the bottle was not there.

Robert poked around in the fridge, thinking that someone had just moved it. Or maybe used a little bit, and put it back by mistake on their own shelf; that wouldn’t be the end of the world. When his bottle of media still failed to turn up, Robert looked all over: in the water bath, on the shelves in the tissue culture room, in the trash containers. Nothing.

Robert went around the lab, asking everyone if they had seen his media. There were only a handful of people around as it was still early, but no one knew anything.

“Just make up a new bottle,” said Kelly. “It shouldn’t take that long. At least your cells are still there this time.”

At this point, Robert had spent a good part of the morning trying to track down his bottle of media. He still had plenty of time to make more media and run his experiment, but he was feeling exhausted. In particular, the emotional strain of suspicion, counteracted by his own attempt to contain that suspicion, was wearing him down. He decided to take a short break and get some coffee before starting over again.

The coffee room was on the first floor, down three winding flights of stairs. Usually it was crowded but when Robert arrived the room was empty. A full pot of coffee sat in the coffeemaker, still warm. Robert swirled the pot, transfixed for a moment by the translucent brown-ness of the coffee, like mud diluted very finely.

While scanning the counter for a mug, Robert’s peripheral vision caught a movement from the direction of the door. He looked up; it was a young woman he had never seen before. Her hair flowed down the back of her neck like hazelnut chocolate, swirling from side to side as she moved, held in place at the top of her head with a red scarf. Robert’s eyes followed her as she reached the far side of the room, turned, and sat in the corner on the couch. His eyes stayed glued to her, but she stared straight ahead, seeming not to notice him, and began flipping through some papers on the coffee table in front of her.

Briefly, she looked up and they locked eyes. Hers were hazel and round, like two saucers needing a refill of espresso. She looked back down at her journal article. Robert stood there, still holding the coffee pot, still without a mug, cells and media temporarily forgotten, thinking about whether he should say something. She looked up and saw him staring at her; he turned his eyes away; she tilted her head back down. Damn! He thought, feeling stupid. He turned back to the counter, found the clean mugs, picked one that said “Drink Me!” and began filling it.

When he looked up again, she was gone. He cursed himself again, thinking she must have slipped by when he was facing the counter. It made some sense to him: her body, so lithe and soundless, almost not present. Translucent, she had seemed to him, like a porcelain doll that might break. He shook himself; added sugar to his coffee. He took a sip and as the warm liquid drizzled down his throat he felt the caffeine oozing through his inner organs and out to his limbs. He shook himself again, wondering what had come over him suddenly, why he was so overtaken by a woman he had never seen before, one who was not even really that attractive. Or was she? Robert decided it had just been his bleary pre-coffee state. She wasn’t even his type. He had been so upset about his recent lab mishaps that he was daydreaming, perhaps had even dreamt her up. How had she disappeared so quickly? In any case, Robert told himself, if she worked in this department, he was bound to see her again.

Energized, Robert went back to lab. He got a new bottle of media from the lab’s cold room, thawed and added the necessary supplements, filtered the bottle. This time, he was careful to label it with his full name and the date, in larger letters than usual, using the dry erase marker. He wrote on both sides of the bottle, as well as on the lid. Not taking any chances. Robert continued with his experiment – collecting cell lysates and passaging the remainder of the cells – and was pleased that he was still able to finish before dinnertime. He put the leftover media on his shelf in the tissue culture room fridge, threw the empty tubes and plates into the biohazard trash. Removed his gloves and washed his hands, letting the cold water from the sink drip into his empty palms. Still thinking, in spite of himself, about the woman from the coffee room.

To be continued…

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About the author

Deborah Flusberg has a PhD in cancer cell biology and now works in biotech in Cambridge, MA, USA. Her research has included studies of cell life and death and she is interested in the human elements that motivate scientific research and the interplay between art and science. She has been dabbling in fiction writing since she can remember; this is her first LabLit.com publication.