Wednesday, July 2, 2014

the altruism of biotechnology + aseptic requiem

Woven catgut sutures on 3D printed ABS loom, now enculturated with 3T3 cells. Loom & needle printed at Alternate Anatomies Lab with Stelarc.
It struck me last night (as I was attempting to hand-weave the above miniature cloth of surgical suture thread--which is, it turns out, decellularized cow/sheep gut--within a sterile laminar flow hood, touching as little as possible, gloved and gowned and sprayed down repeatedly with ethanol, and using sterilized tweezers to attempt to hold and manipulate my miniature tools to do the weaving), the deep contradictory nature of employing a handcraft technique to a laboratory process. Or is it?

Surgeons still stitch their patients up by hand as far as I know. Skilled textile and flesh artists they are! The below diagram of minor surgical stitches is something I find fascinating, particularly the vertical mattress stitch.


Anyhow, I am doing biological weaving - something that hasn't yet been done in either the scientific or textile arts worlds, to my knowledge. Perhaps the creation of a diagram is warranted. Using suture thread of gut is perfect for my work with the cell cultures because it naturally deteriorates in about 90 days, long enough for the cells to take over the structure. The gut thread maintains its original tensile strength for approximately a week. It will be interesting to watch my weaving degrade and change as the cells work away at forming colonies on it.

3T3s are my best first test cells to use for the woven scaffolding because they grow like weeds. 3T3s are connective tissue cells. I now have five healthy flasks of 3T3s that I grew myself, after receiving a gift flask from a fellow artist-researcher here at SymbioticA, Soichiro Mihara from Japan. Soichiro afffectionately refers to us as 'cellmates' now. Soichiro has been treating his 3T3 cultures with low levels of gamma irradiation in order to disable their genetic ability to reproduce. I went with Soichiro last week to the Princess Margaret Hospital for Children, where we used the gamma irradiation tank. My purpose there was to sterilize my miniature looms and tools with very large doses of gamma irradiation (>25kGy). This was an experimental method for sterilizing my looms because I can't sterilize them by autoclaving--the 120˚C steam would warp the looms--and I can't sterilize them by UV radiation because the ABS plastic they're printed of is initially UV cured when it comes out of the printer, so futher UV radiation will deteriorate the material. Alas, what I discovered when attempting to sterilize using gamma irradiation is that the irradiation vault we had access to is set permanently to a low level of radiation and in order to irradiate my items to the level required for sterilization, they would have to be irradiated for 11.5 hours continuously. I just couldn't use the machine for that long. So, failed effort. I settled on ethanol sterilization, even though it too degrades the material that the looms are made of, but not to the extent that it will ruin them. So, it is the best option out of my available options. The absolute best method would be cold gas sterilization, but I don't have access to those facilities.

Here are some images of 3T3s from the digital microscope:


I named my first 3T3 culture after my grandmother, one of my 'cell parent' donors from my crowd funding campaign.


Everything is pink in the flasks and dishes because the cell media (food) has a pH indicator added to it so that it is easy to tell if the culture is unhealthy (too acidic with ammonia or too basic) or not. I am pleased, since I really like the pink as an aesthetic component.

I've thawed and plated my osteoblasts that I received from Audrey Chan, the PhD student and researcher at the QEII Hospital, and they are healthy and happy, I'm thrilled to report. This is a pinnacle moment, as just last week I euthanized every miserable, dying and otherwise useless cell culture I had in the incubator. This includes the historical SymbioticA pig wings cells, proving that a moment in history cannot be brought back to life. Essentially, I started from scratch again last week, with 3T3s as my saving grace. I was extremely nervous thawing the osteoblasts, because they are my last shot at being able to grow bone while I'm here--my last access to bone cells, really, since my primary sourcing directly from bone did not pan out in the end. Anyway, my osteoblasts are thriving in regular D-10 media, with a new batch of bone soup on the front burner (not literally but the metaphor is nice).

When euthanizing my last batch of cultures, I took a moment--the 30 seconds or so that it takes for the bleach to do it's killing job--to thank the cultures for all they had taught me in the last two months. And then, I wrote a new lab protocol for 'Aseptic Requiem for Cell Cultures'. The protocol is below:


The requiem (or service) for cell cultures is appropriately brief and timed and convenient with the rest of process. This is done purposely to highlight the level of emotional distance or complete lack of sentimentality when dealing with life in a laboratory environment. The idea of an aseptic requiem is ludicrous. I'm also toying with that false dichotomy between science and religion by calling it a requiem, which is essentially a Catholic mass for the dead. Anyway, I did say a nondenominational prayer of thanks to my cell cultures before disposing of them because for me, acknowledging and reflecting on the life that I'm working with is important for maintaining my sense of humanity.

I also thawed my extra C2C12s that I had initially grown and they are doing their thing in their flasks. The previous flasks of C2C12s I had essentially starved to death because I had been feeding them differentiation medium, which is only 2% fetal calf serum versus the 20% they thrive on. Differentiation medium turns the C2C12s from single nuclei cells into myotubes, or joined together, multiple-nuclei cells. I probably should have been feeding them regular media (10%) after they differentiated but Ionat was traveling abroad for three weeks and I had no guide. Interestingly enough, Ionat just returned from the Viconti lab in Boston, Mass. and learned a new protocol that is similar to my project with using silk (only it's not woven). My silk is now a beautiful milk chocolate brown after dyeing it with my sterile-filtered, autoclaved eucalyptus bark dye. Not only is it beautiful, but it is now slightly antibacterial from the eucalyptus, which can only be a good thing in a cell culture.


One last consideration--something I've pondered in the last few days: the donors who became 'cell parents' during my crowd funding campaign are by extension, 'cell donors' even though the cells have not come from their own bodies. They are surrogate cell donors, of C2C12s, 3T3s and now mouse osteoblasts. I will attempt to grow new embodiments from this disembodied life. The altruism is manifold. What about altruism in science? 
In watching a documentary about Ayn Rand and her (I think, sociopathic and reactionary against communism) philosophy centred on Individualism, as well as its link to the development of technology, the crash of the financial world, and essentially the general degradation of society one might argue, I think there is a place in the world for altruism, and perhaps in biotechnology. More to unravel for next time. 

No comments:

Post a Comment