Science, Working, and Working Out

Hi all!  I hope that you're doing well.  But enough about you, this is my blog.  Thus, we talk about ME!  Yay me.  So I don't have my latest video ready yet (a video game review this time; pretty major FPS).  Long story short I've been working on getting the technology to record gameplay as well as upgrading my video editing software (for whatever little it may be worth).  Moving on....

Working out.  Backwards from the title?  Yes.  Do I care?  No.  Anyway, Christine and I have been doing the Insanity program for two weeks now and it's pretty rough.  We're both doing better, though, getting stronger, losing some fat, and I'm fairly certain I have an ab.  It's pretty awesome, if I say so myself.  Which I do.  If you've wondered about it or heard and don't know much, it is pretty worthwhile.  We like it, anyway.

Science and work!  Work and science.  Hmmm.... A thought occurred to me that most people I know aren't aware of what I'm studying.  If you're curious, read on.  If not, well..... watch the videos.  They're pretty cool.  And subscribe on YouTube.  Now let's see... right!  My science!  I'm doing work with mouse Embryonic Stem (ES) cells.  These were popularized around 2000 with their massive potential for therapeutics, etc., but underlying ethical issues of destroying fetuses to harvest the cells.  Enter line restrictions  protests, vandalism, etc.  Regardless, they do have substantial potential for therapies with diseases causing cell loss (Alzheimer's, Leukemia, Parkinson's, etc.) as they have the ability to turn into any cell type in the body, from the brain to the toe.  Though I can't think of a toe degeneration disease off hand.....  Hahahaha!  Can't think of a TOE off HAND.  I crack myself up.  Anyway, these so-called pluripotent ('many powers' in latin; in this case can turn to many adult cell types) cells are taken from the inner cell mass (see below) of a developing embryo and can be kept in culture indefinitely.

Establishment of pluripotent Embryonic Stem (ES) cell lines

Again, there is a great deal of controversy with this, namely amongst religious groups (see science education and evolution vs ID videos for some more controversies).  In 2006, it was discovered that you could take an adult cell (liver, skin, brain, blood, etc.) and, by forcing it to express specific transcription factors, change it back to the state of pluripotency, thus birthing the induced pluripotent stem (iPS) cell.  This was a major breakthrough in the technology, just recently indicated by a Nobel Prize win.  Instead of trying to get these dissident groups to go along with using ES, we found a way to get around the controversy entirely.  Not necessarily easy, but it certainly worked.

Examples of Reprogramming (bringing cells back to pluripotency) and some of the methods used.

Before we start randomly injecting these, or other, stem cells randomly, though, we need a few things.  Namely, we don't know a great deal about how stem cells work or the processes that change them to new cell types.  Don't get me wrong, we know plenty about it, but not enough to start with therapies.  My project is based around a specific aspect of this: how the cells deal with mutations.

Mutations are permanent changes in the DNA sequence of cells.  Frankly, ES cells don't mutate very much.  Data has shown that they mutate at about half as frequently as adult cells and about a quarter as much as cells made in the lab using ES (for example, making a brain cell from ES in a culture dish).  My project is focused on trying to understand why this happens and, perhaps, eventually be able to maintain this level of genetic integrity (as it's called in the field) when the cells are becoming whatever they need to be for the therapy.  I'm specifically looking at the major pluripotency genes; the ones that keep a pluripotent cell pluripotent, and seeing if they have some relationship to genetic integrity genes.  There are two major ways mutations are fixed: DNA repair and Cell Death.  DNA repair fixes smaller issues (single or a few bases missing, mismatched, etc.) while cells trigger apoptosis (a major death pathway) is there is major damage such as too few/many chromosomes, large scale damage, etc.  What I've found by looking at gene expression and control data is that there is a great deal of interaction between pluripotency and these integrity genes.

Cell death genes (right) under direct control of the three major pluripotency genes (left)

If we add in additional transcription factors, the number of controlled genes increases significantly.

I'm also going to be validating this data in our own ES cell lines, ensuring that the binding is occurring and the expression levels are the same as this data shows.  In addition, I'll eventually be getting rid of the plutipotency factors from our cells and seeing how both the integrity gene expression and mutation frequency are affected.  To make a long story short, we use a transgenic mouse that gives us the frequency of mutation by a special reporter gene.  It's pretty awesome.  As a quick note for you, as well, here's a special picture from a couple of days ago:

This is an image of my cells in culture.  The larger white circles/ovals are the cell colonies while the longer, individual cells are a cell type we use to grow the ES on, known commonly as feeders (appropriately, as they take in the media we give and spit out the media the ES cells use).  Some of the smaller bunches of cells are the differentiating ES (those turing to adult cell types), particularly the ones with distinct nuclei (a lot of white space and a small dot).  And if you're curious, this picture is of the cells at 40X actual size.

I hope you liked my little overview of my project.  If you're feeling extra nerdy, feel free to ask about it.  If there's one thing I enjoy it's talking about science, and more specifically my work.  Either way, I'll see you next time!