doc w/ Pen

The journalist in me is coming out. And so is the future physician. “How can this be?” you might ask. Well, read on.

My love of news and my understanding of the news business have come together with my passion for medicine to inspire an idea: a weekly column, written by me and presented here on this blog, about current events in medicine and related subject areas (such as medicine and politics, etc.). I call it “Health in the Headlines,” as indicated by this blog post’s title. This feature may be, in part, what we referred to at my old newspaper job as a “news roundup.” As in, it may contain summaries of news stories that I have found throughout the week from various sources. Another possibility is my choosing a controversial issue that has been in the headlines steadily and drawing from several weeks, even months, of stories for information. This feature may also contain aspects of an editorial. Or if time is short, perhaps I’ll just post links to a handful of really great stories or podcasts and leave it at that. Oh, yes–pictures are great too. And they fill a lot of space. (This is one thing you learn very early as a journalist.)

The point is, I am going into this “feature” with flexibility. I do not want to hem myself in by saying I will ALWAYS do this, or I will NEVER do that. Nope, I want to do what I feel like doing, when I feel like it. Because after all, I want to have fun with this. But I promise that it I will try my best (as I do will all my blog posts) to make it interesting.

One more thing. You might be wondering why I’m trying to keep track of current events in health, medicine, research, etc. One reason is that when I’m interested in something (in this case, medicine), I want to learn more about it. Obviously, I will learn a great deal in school. But what you learn from the New York Times is very different. First of all, you hear from real people–not academic textbooks that are dry and technical. From those “real” voices, you get a sense of what other physicians are doing and how medicine is moving forward (or not). You get a sense of what interests you and what doesn’t. You start to develop passions and opinions and your own sense of morals and ethics.

Another reason I am doing this is to form good habits for the future. When I am a doctor, it will be necessary to keep up with this type of information (especially that pertinent to my specialty). If I start now to keep myself in the loop, it will be but a small step when I have to keep up with medical journals.

Furthermore, it’s simply good–no, necessary–to be prepared with both background and contemporary knowledge of your field. Not only will it come in handy when I am a physician (knowing the most recent treatments for X, Y, and Z), but it is also useful now. For example, when I shadow a physician, knowing something about his or her specialty allows me to ask intelligent questions (which you should do not for the brownie points but so you can actually learn something). This in turn yields specific answers. And at the dreaded medical school interview, there will also be current events questions. Cramming for that interview–which is what most people do–is a horrible idea, just as cramming for anything is a horrible idea. But if you’ve followed what’s been happening in medicine all along, those questions will be cake. (And I very much like cake.)

So this is the idea. We’ll see what happens. Maybe I’ll try it and it’ll fizzle out, who knows. For the time being, consider yourselves journalistic guinea pigs. Hope you enjoy the ride.

Note: I will post the first in this series later today (Friday, June 25).

Practice makes perfect.

I hate that phrase. In fact, I wish they (I’m not sure who “they” is, though) would strike it from the annals of common English phrases.

As an eldest child, and a dyed-in-the-polyester (I prefer vintage to chic) perfectionist, I have been working, for the last several years, to deconstruct my desire for perfection. Because absolute perfection, as pictured in my perfectionist brain, is absolutely impossible. Striving for the impossible is pretty pointless. And, as I have discovered, exercises in futility are infinitely frustrating.

So, here is my replacement phrase:

Practice makes improvement.

Or, as the Beatles put it:

Getting better all the time … 

And this morning, as I was discussing my recent work at the lab with my husband, Geoff, I realized something. I am practicing. I am improving. I am getting better all the time.

Here are a few of my observations:

• My speed and technique have improved dramatically.

When I first started at the lab, I had to start from scratch: learning to pipette. My hands were shaky with nervousness (and sometimes over-caffeination), and I often contaminated the tip of the pipette by touching what I wasn’t supposed to–the outside of a flask, the countertop, you name it. I just didn’t have the mentality of keeping things completely sterile. Of course, I would realize my mistake right after doing it, but by then it was too late and I had to discard the pipette (and what was in it) and tear open a new one. This wasted not only money and supples, but also time.

These days, I rarely contaminate the pipettes. I move more quickly from bottle of solution to test tube or flask. My hands are more steady. I have better aim when releasing liquid (again, less waste, more accuracy, and time saving). Olga even trusts me to “passage” the cells now–divide them from, say, four plates (dishes) into eight through a series of washes and “trypsinization” (which dissolves the cells’ hold on the plastic plates and allows them to be sucked up and released into new plates–a very delicate procedure). This process has to be done very quickly or the cells can be severely damaged, even destroyed. I’m fast enough–and accurate enough–to do this now.

• I am more autonomous.

These days, contrary to my first few weeks in the lab, Olga will give me an assignment and pretty much leave me to it. Of course, if I have questions or a problem, she is available to help, but she has the confidence–and the expectation–that I will remember the techniques, processes, and steps that I have learned and can (and will) repeat them succesfully and in the proper order.

Thankfully, Olga is an amazing teacher and explains things extremely well, so that by the time she expects me to do them on my own, I am more than prepared to do so.

The fact that I am more autonomous forces me to rely on myself more, and learn the techniques for myself, rather than rely on Olga to re-explain them every time. This dynamic also allows Olga and me to accomplish more because we can both be working on different things at the same time. Even though I work more slowly than Olga, slow work is better than no work. As she put it the other day, “If you hadn’t done this [set up a reaction], I wouldn’t have had time to do it at all.” That was the best compliment she could have given me.

• I am more self-confident in my work.

Self-confidence has never been one of my strong points. Or so I thought.

When I was a reporter, I remember walking away from finishing the newspaper on deadline day thinking to myself, “Did I spell that commissioner’s name right?” or “Did I put the right date in for that park district event?” or “Did I write a caption for that photo of the dog park?” Heart pounding, sweating, a total nervous wreck as I walked the few blocks to my car, fighting the urge to turn back to check on whether I had done those things or not. I knew that I had, but I doubted myself anyway. I was sure I had forgotten something, so I mentally went through just about every possible mistake I could have made trying to find my error (or errors). With my lab work, I have very few such thoughts. And if one starts to creep in, I immediately squash it like the ugly, infectious cockroach that it is. I trust in my precision, my accuracy, my work ethic, my knowledge, my understanding, and my ability to follow directions. One thing that has really helped me is to develop a process for myself when I work on a task.

For example, if I am making multiple “master mixes” for a polymerase chain reaction (also called PCR–a process that creates a bunch of DNA from a very small sample), I create a checklist for the “ingredients,” along with the microliters needed of each. First, I will load the magnesium chlorite into each test tube, then put the MgCl off to the side so I know it has been added. Next I will load the “super clean water,” check it off, and put it off to the side, and so on. This way I don’t duplicate any ingredients, and I know when everything has been done (and in the proper amount). This, of course, also contributes to speed and efficiency–which never hurts. For times when those “cockroaches” scuttle up, I have, already prepared, a list of battle phrases (or cans of RAID, if you like) to use against them.

An example: “I acknowledge that I am concerned about whether I added the cDNA to the master mix, but I have checked it off on my list, and the tube has been put off to the side. So even though I don’t specifically remember adding it, I know that I did because I followed my process. As a result, there is no need to be worried about this.” Maybe it sounds hokey, but using these phrases, for the few times I have to deal with worrisome and negative thoughts, really makes a difference.

I have more self-confidence now to begin with, and when I struggle, I have another “tool” in my mental “toolbox” to help me.

• I better understand the science involved in what we (and other scientists) do.

On May 11, my first day at the lab, I understood … um … maybe 10 percent of what Olga was telling me. OK, perhaps a little more, but it was bits and pieces, not a comprehensive picture. So I spent a lot of time being really confused and doing things but not really understanding why.

Please note — this was not Olga’s fault; she explained (and continues to explain) things very well. I simply did not have the background, nor the scientific vocabulary, to comprehend even the most elementary of explanations.

Boy, how things have changed. And I especially noticed it this week. On Wednesday, Olga had me set up the PCR reaction (see explanation of PCR in observation No. 3). Before I started pipetting, she gave me the background on what exactly we were going to be looking for in the end (with the gel electrophoresis). I took very careful notes, both so I could set up the reaction properly and so I could remember the details of what I had done. The same buzz words came up as had come up before: transcription factors, differentiation markers, fetal organoids, endogenous control. This time, though, I actually knew what she meant. In fact, I knew enough to ask intelligent (and genuine–not pedantic) questions about what she was telling me! It was a true victory.

I’m not much of a hymn-singer, but I totally got what John Newton meant in “Amazing Grace,” though in a scientific, rather than religious, context: “I once was lost, but now am found / was blind but now I see.”

The second example of this “understanding” phenomenon came at yesterday’s (Thursday’s) weekly afternoon lab meeting. Each week, the usual procedure is that one of the lab’s scientists presents his or her recent work for discussion. This week however, we did something a little different. Lab Director Rich Minshall and one of the PhD candidates, Aaron, picked out a research paper for us to read and discuss.

When I read the title early Thursday morning, I groaned. My eyes glazed over, anticipatorially, sure that the paper was going to be a total drag (i.e., completely incomprehensible to me). It was indeed just the title that gave me this impression: “Transcellular migration of leukocytes is mediated by the endothelial lateral border recycling compartment.” *GROAN* indeed.

But I started reading anyway — that damn Puritan work ethic again. And to my surprise, I was understanding things. Sure, I had to reread parts, sometimes multiple times, and look up certain concepts on the Web, but I was getting it. In fact, I was getting into it — I even started highlighting the text onscreen (I’d never done that before; it was pretty cool) so I could remember and review the highlights and main conclusions / points of the paper before our 4 p.m. meeting.

I didn’t really understand the visuals that accompanied the paper, but that was OK. I understood the ideas. And at the lab meeting, I actually contributed — for the first time. Not on the level of the Phd’ers, but who would expect that? The fact remains, I contributed, because I was able to understand what was going on–something that had never happened before.

I felt kind of like a surfer who, for weeks, had been trying to catch a wave, only to be knocked down to the ocean floor. But one day, she manages to hold her balance, and hold the wave’s crest, and ride it through. Pretty cool.

Surfing science … I kind of like that.

Sounds impossible, right? That anyone would get confused about whether they were at home or in a scientific laboratory? Well …

OK. I’m just kidding. I don’t really get confused about the two. But I have noticed two interesting trends: 1) There is a lot of my “home” in the lab. 2) There is a lot of “lab” in my home.

Let me explain.

First: If you entered the E420 lab where I work, you would see all kinds of foreign-looking equipment that has virtually no place except in science. Test tubes, pipettes, fume hoods, microscopes, crazy chemicals. But mixed in with the foreign is a good bit of the familiar. And when I say “familiar” I mean “household.” As in, tin foil (used to cover beakers and test tubes), plastic wrap (used to cover gel when taking it to the dark room to photograph), masking tape (for labels). There are also – I kid you not – regular, everyday, conventional freezers (the upright kind), refrigerators, and microwaves. I used the latter just today to make agarose gel for gel electrophoresis. (I would NOT recommend using this microwave to also warm up your coffee. Just a thought.)

Second: Working in the lab becomes a mindset. It begins to pervade your thinking in a way that you can’t simply shut off when you flip the lab’s lightswitch at the end of the day. For instance, I was at Starbucks the other day getting coffee. I went over to the little condiment stand to “dress” my coffee with sugar and cream. I was about to set my lid down on the countertop when a red warning light went off in my brain: CONTAMINATION ALERT! Rather than just set the lid down, I quickly picked up a clean napkin and set the lid on that. Whew. Crisis averted. In a similar vein, I was at home filling a water bottle. I unscrewed the lid and was about to put it next to the sink, thread side down, when that same warning went off in my head: CONTAMINATION ALERT! I quickly turned the lid so that it was the top of the lid that touched the countertop, not the bottom – thus avoiding possible (or in my house, probable) countertop contamination.

Crazy, I know. But when you’re working with very fussy fibroblasts and epithelial cells that whine and scream (translation: DIE) when you put the lid down the wrong way, or brush your fingers up against the top edge of the container, or pass your (gloved) hand over the open container (obviously, dropping skin cells and other yucky things into the culture dish that the cells won’t like), you become a little paranoid about contamination. When touching *ANYTHING* with a sterile pipette tip means it’s no longer sterile and you must throw it away, you start to look at “inappropriate contact” of objects as a serious problem. And this mindset, as I said, is very difficult to quiet, because it is necessary to keep it at full blast all day long. Relax for a minute, and you risk ruining a reaction (or several).

Does this mean, at the least, that I’ll become a better housekeeper?

“This is a marathon, not a sprint.”

That’s probably the most often-quoted line on www.OldPreMeds.org, an online forum dedicated to so-called “non-traditional” pre-med students such as myself–students who want to go to medical school after time off from college, after another career, after having children … you name it.

Comparing the journey to, and through, medical school is quite appropriate. First of all, it’s long. Especially if (like me) you have a year or two of pre-requisites to take. It’s also grueling. Extended work and study hours, lack of sleep, stacks of material to learn. But you learn so much about yourself along the way, and at the end, have accomoplished something incredible. To get there, though, you have to pace yourself, rehydrate along the way, and have trained your ass off before you even started the race.

Another key to finishing this “marathon” is surrounding yourself with supportive people. Family and friends are indispensable, of course, but you also need people who are going through the same journey and can relate, step by step, to what you’re going through. That’s where OldPreMeds comes in, at least for me.

I started off as a “lurker.” A person who perused the online forum but who rarely, if ever, posted anything. I didn’t believe I had much, if anything, to contribute to the discussion. After all, I hadn’t even started my post-baccalaureate program yet.

And then on June 10 (nearly two weeks ago), Richard Levy, who helps run OldPreMeds, called me out.

I had, at the last minute, decided to attend OldPreMeds’ 10th annual conference, which, luckily for me, was held this year in at a hotel just outside Chicago. (Otherwise, I would not have gone–I wasn’t familiar enough with the organization and didn’t have the money saved.)

It was at the preliminary cocktail session that my status as a lurker was revealed. The rest of the conference, however, was so inspiring that not only have I ceased to be a lurker on the online forum (I’m up to 46 posts so far!), but I’ve started a Chicago affiliate of OldPreMeds (OPM-Chicago). My goal is to continue the face-to-face interactions, networking, and support that was so helpful at the conference.

So what was so incredible about two and a half days spent in a Holiday Inn? PLENTY! Here are some of my highlights, in no particular order.

– Getting a breakdown of the application process. (from Judy Colwell’s “The Nuts and Bolts of Applyint to Medical School As a Nontraditional Applicant). I had no idea that this is a 15-month process, starting in the winter with contacting your reccomendation writers and ending as late as the next summer with final decisions from med school. But now I know what to do in between — when to request transcripts, when to take the MCAT, all that good stuff.

– Being the ADCOMS. “ADCOM” is the acronym for “admissions committee,” and in one of our workshops, we got to play the role of the ADCOMS. The presenter brought in real applications (with the names, addresses, etc. blacked out to protect privacy) and had us work in groups of five or six. Our mission? To decide whether a particular applicant would receive an interview invitation. It was eye-opening to see what ADCOMS (should and do) look for, beyond GPA and MCAT scores–motivation, initiative, community service, clinical exposure, writing skills, an updward trend in grades, a depth and breadth of life experiences, etc. I now have a better idea of how I will structure my own application (when that time comes).

– Professionalism and admissions. Duh, right? Don’t wear a tube top to your interview. But no, wait–there’s a lot more to “professionalism” in admissions than how you dress. Examples: 1) no questionable photos/posts on Facebook. 2) Get to know the admissions reps where you intend to apply to school (and summer is the best time to do this, because they are the least busy). 3) Visit the campuses where you intend to apply–schools may look very different on paper than in person.

– Richard Levy’s “10 Things Every Non-Traditional Applicant Should Know,” such as: Take a Breath; Don’t Bite Off More Than You Can Chew; Make The MCAT Your Friend; Don’t Risk Bad Grades By Doing Too Much; etc. Common sense things that we (or at least, I) often forget in the midst of stressful times.

– Networking. I got phone numbers and e-mail addresses from fellow OPMs, as well as M.D.s and admissions people who may be willing/able to help me out (including a professor at the University of Illinois at Urbana-Champaign, where I want to apply for school). Note: networking happened at the conference, as well as into the wee hours of the night …

I left the conference walking on clouds. Because for the first time, I really felt like I wasn’t alone.

Note: This post was written 5/29/2010, while I was on a plane on my way to Hawaii. I had no Internet access on the Big Island, so I am just now posting this.

In no time at all, I launch my official pre-med journey.

T-minus 3 months … 

Yes, that “3 months” refers to the beginning of school. Hard to believe that in a short 90 days or so, I’ll be a student again, after a 7-year stint as a grown-up.

So much feels the same, and yet, so much has changed.

I have yet to finalize my course load — so far I’m registered for Chemistry, Biology, Calculus, and a 1-hour “Topics in Medicine” class (it’s a requirement for me). I’m also trying to get into Physics, which my advisor has assured me will be a non-issue. That would put me a 17 hours – a pretty heavy load for someone who hasn’t been a student in nearly a decade. So I’m in the middle of a debate with myself: do I take all of these classes? Take Physics next year instead? But I’ve heard taking Physics with Organic Chemistry (a so-called “weed-out” course I have to take in the 2011-2012 school year) is a nightmare. Then there’s the issue of my wanting to take as many of the “fun” upper-level electives as I can, which means getting the basic pre-requisites out of the way ASAP. *sigh*

This reminds me of when I was preparing to start my freshman year in high school. The debate then was whether to take a 2-hour accelerated science course, or just take the 1-hour basic (in my mind: “dummy”) science course. I remember being at my grandparents’ cabin in Colorado, sleeping on the pull-out bed and obsessing over this dilemma. (In the end, I took the easier class because the harder one just didn’t seem worth it; plus it didn’t fit in my schedule very well. It turned out to be a good decision.)

However, this time around, I am not obsessing. Actually, I am very calm. My approach? Sign up for all the classes, start off the semester, and see how things go. If the classes are too much, I’ll drop one of them before the “drop” date and no one will be the wiser.

In these 15 intervening years, between the ages of 13 and 28, I have learned to have faith in myself, and in my decision-making abilities. I am more flexible. I have learned to handle change. I believe things will work out, and work out well. The thought of the unknown, of uncertainty, does not leave me panic-stricken.

Some things stay the same. Thankfully, other things do change.

Note: This post was written 5/29/2010, while I was on a plane on my way to Hawaii. I had no Internet access on the Big Island, so I am just now posting this.

Large “Red Eye” (coffee plus a shot of espresso), room for cream. That’s my usual order at Descartes Coffee, the coffee stand in the University of Illinois at Chicago Student Center. Just across Wolcott Street from the Medical Sciences Building where the lab is located, the Student Center is a hopping spot at lunchtime for medical students and other UIC staff and faculty. I don’t usually talk to anyone there – other than Eva, who staffs the Descartes stand – I just watch, and listen.

You can tell a lot about a person at UIC by what he or she wears. White coats, for example, inside the lab building suggest a researcher or PhD student. The same coats worn outside the lab usually denote medical students (short coats), residents (medium length coats) or attendings (long coats). A stethoscope around someone’s neck is a dead giveaway — definitely the medical variety.

I visually size up the length of someone’s coat, try to catch what his or her sleeve patch says (“House Staff,” for example, or “Medical Student”). Without drawing attention to myself, I try to lean in to hear what the people at the next table are talking about — an impossible gross anatomy exam? A grueling surgical rotation? A demanding attending?

People often complain over lunch (which, in the Student Center, usually consists of a Subway sandwich), griping about this or that to fellow students or colleagues. But I soak it in. I dream of the day when I will be in those shoes, groaning about that exam, rotation, or attending. I probably won’t think so at the time, but from where I am now, being able to complain about such things seems like a privilege.

OK, maybe that sounds stupid, and Pollyannish. Although really, when it comes down to it, I’m not exactly your sunny optimist type. But when you think about it, just getting into medical school is a long haul. And once I get there — and I will get there — I do not plan to take it for granted.

Catenin … adhesion junctions … cadherin … p120 … tyrosine … kaiso …

I stared blankly at Dr. David Wei, who was presenting the latest research on, well, um, I think it was about the role of a protein called p120 in the “adhesion” of cells – their ability to stick to one another. Yeah. I’m pretty sure that’s what it was about.

Olga had invited me to the weekly Thursday lab meeting. I was thrilled to be included. Rich Minshall (the lab director) introduced me to the other half-dozen or so people I hadn’t met yet, and asked me for a summary on what I’d learned so far. Everyone seemed impressed with the depth and breadth of what I’d accomplished in a mere three weeks. So far so good.

And then David began his presentation. Nothing against David – he’s an incredibly talented, intelligent anesthesiologist and budding research scientist. But he could have been speaking Greek as far as I was concerned. And the slides he was showing looked like hieroglyphics to me. I simply don’t know the language of laboratory science.

Slowly, as the 45-minute presentation went on, I began to catch on – a few words here, a concept there. By the end, I had a vague idea of his main idea. And I had picked up some new words along the way.

I’ve been in foreign language contexts before. In college, I studied abroad in South America (although I am nearly fluent in Spanish, so that isn’t a particularly good example). A better example: In high school, I went to China for a school trip. The words, spoken and written, all around me bore absolutely no resemblance to the language I had grown up with. It was humbling. To have to ask lots of questions, to have to learn from the ground up, to have a hard time saying “hello” because of the use of intonations (vocal inflections), which we don’t have in English.

My dad has told me over and over that the first two years of medical school are basically spent learning a new language: the language of medicine. I got a small taste of that Thursday. A flood of technical language, technical diagrams, the expectation of rapid absorption and understanding of new material. There will be no coddling, no hand-holding, no “let-me-spell-this-out-for-you.” Catch on, and catch on quickly, or you fall behind every more quickly.

Fortunately, I have a gift for languages. When it comes to grammar – whether it’s Spanish verb conjugation used in conversation, or Latin prefixes and suffixes used in medical terminology, I am quick to grasp the concepts. I also know how to memorize: vocabulary (words and phrases), linguistic rules, and exceptions to those rules. This is how I breezed through high school and college Spanish, and through a semester in Chile living with a Spanish-speaking family. This is also how I will learn to communicate in medi-speak.

And one of the greatest things about working in the lab right now is that it is proving to be sort of an immersion experience, which is, really, the best way to learn a language. I am exposed to terms, concepts, processes, substances, etc. that I have never encountered before. I hear about them over and over. Sometimes someone will tell me, specifically, what a particular thing means or does. Other times, I have to figure it out on my own, over time and through numerous exposures in different contexts.

Basically, it’s the Montessori method: experiential-based, curiosity-led learning at one’s own pace. And it’s joyous.

Lung fibroblasts (pictured above) produce the “food” our lung epithelium cells need to survive, as I learned this week.

A few days ago, Olga told me she’d thrown out a flask of fibroblast cells. What?! I couldn’t believe my ears. Didn’t we need those cells for … um … something?

Every day, I learn new techniques. But sometimes I get caught up in the “doing” and miss the bigger picture. Monday, though, I got some clarity. The “doing” and the “bigger picture” came together.

Olga and I are taking care of two types of mouse lung cells: fibroblasts and epithelium. Up until Monday afternoon, I couldn’t keep them straight.

Monday morning — before the clarity — Olga had me work with 10 plates (round plastic dishes) of fibroblasts. For five of the plates, Olga instructed me to aspirate (remove) the high-calcium media (liquid they grow in) and pipette in low-calcium media. Simple enough. Aspiration, pipetting, easy stuff (which feels really good to say!). I wasn’t quite sure why I was replacing high-calcium media with low-calcium media, but Olga was busy setting up another experiment so I went ahead with her instructions.

Once I was done with that, Olga told me that we were going to “passage,” or split, the other five plates into 10 plates to give the cells more room to proliferate. Basically, we wanted them to grow more, and they needed space to do that. That’s all well and good, but I wasn’t quite sure why we needed so many fibroblasts … we already had a lot, and had more in the freezer; why keep so many in the incubator? A question for later, I decided.

The first step in passaging the cells was aspirating their media. Then I washed the plates out with PBS, a sterile buffered saline solution, to remove any traces of serum from the plates (serum is one of the ingredients in the media). After aspirating the PBS, I added a solution of trypsin and EDTA to the plates, one by one, to detach the cells from the bottom of the plastic plates. Trypsin is an enzyme found in the digestive tract, and it literally “digests” the proteins that help the cells stick to the bottom of the plates. Olga and I then waited a few short minutes for the cells to detach, then pipetted in more media to inhibit the trypsin (to keep it from completely breaking down the cells).

I then sucked up the cells with a pipette, put them in a centrifuge tube, and ran the centrifuge machine for a few minutes. The result was a tube with a “supernatant” – layer of light pink liquid on the top — and a “pellet” — solid collection of cells on the bottom. I aspirated the supernatant, leaving the pellet at the bottom (this is a bit tricky – you have to be REALLY careful not to suck up the pellet, because then you’ve lost all your cells and you’re screwed). Next step was to “resuspend” the cells in new media and divide them into 10 plates (remember, we started with five, so we’ve doubled the number of plates). Because they have more space, the cells will spread out and grow more until they reach “confluence” – saturation.

OK, I get all that. But why did we do all of that to the fibroblasts? And what about the epithelium?

Olga explained: we want to grow as many fibroblasts as possible (hence “passaging” the fibroblasts from five into 10 plates so that they will proliferate more) because fibroblasts produce the “food” that epithelium need to survive. And the epithelium are the cells that we are interested in looking at, from an experimental point of view, but we can’t cultivate them without also growing fibroblasts.

Basically, when you put fibroblasts into low-calcium media (as I did with five of those plates Monday), the fibroblasts produce what are called “growth factors” that epithelium need to live. Olga threw out those flasks of fibroblasts a few days ago because she had gotten what she wanted from them – the growth-factor-filled media to give to the epithelium – and the cells were no longer any good (i.e., capable of producing more growth factors).

Ah-ha. That’s why you ask questions. To discover the “why.” To give your activity context. To provide you with a goal.