joining history + denatured soup

My rat bone cells have been growing, one at a time, slowly, slowly. Here are some new images from the digital microscope last week:

This is possibly a cell dividing.

Little beauties. I'm just in love with them.
Here are some of Paloma's sketches for what she's thinking about with animating the cells. I think she's got sea life on the brain, too.

Paloma Dawkins.

Paloma Dawkins.

Last week I picked up a cryovial of frozen mouse osteoblast cells from PhD student, Audrey Chan at the QEII Medical Centre, School of Surgery, Centre for Orthopaedic Research, UWA. It was an amazing experience, because Audrey took the time to sit down with me and explain her process of primary sourcing bone cells from rodent femora (long bones). I will thaw the cells this week.

Also, I recently thawed some very special cells - the last cryovial of pig osteoblasts, from a project that Oron Catts did 13 years ago, called Pig Wings. So, the cells I thawed and plated (meaning put them in cell media in pitre dishes) were in a deep -80˚ sleep for the last decade and a half, almost.

Once I've successfully cultured these two types of osteoblasts, I will use some for my project and then I will also freeze some down, so that future SymbioticA projects can use them. There is something awe-inspiring about being a part of the continuation of life in this way - it almost makes me like family. I threw out almost all of my first attempts at culturing bone cells from a primary source (from the cow bones I got at the butcher) because nothing was left living in the dishes. So, now I'm down to about 25 culture dishes that I'm maintaining. I had a lab disaster last night that has threatened all of my cultures now, but I'll talk about that in a minute.

I picked up my 3D printed miniature looms yesterday from Stelarc's lab, and it was a delight to see them come out of the wax! Some of the models failed while others turned out perfectly. The problem with the failed objects is that when I scaled them down from human size to cell culture size, the proportions made the objects too thin in areas and they just didn't hold up. Important lesson in designing for 3D printing. I haven't taken any photos of the miniature looms and weaving tools yet because I still need to clean the remainder of the wax support off of them. I haven't quite figured out the best way to do that, but I'll try 100% ethanol. 

I've been developing my own protocols as I go along in all these experiments, and now have a binder stacked with protocols for everything from preparing silk for tissue culturing to preparing eucalyptus for dyeing tissue to feeding cells to microscopy, etc - I've got about 20 lab protocols now, a blend of science and textile methods. Today I boiled little silk skeins in a 2000ml flask in ultrapure H2O and sodium carbonate. I boiled them in fresh solution twice to remove all of the sericin and industrial chemical residue. The first water, after boiling for 2 hours was quite cloudy and yellowed, and the second one was a bit cleaner. Then I dried them overnight in a laminar flow hood (sterile conditions), and they are now double-bagged and sealed in autoclave bags to be autoclaved in 120˚C steam, to make them completely sterile for working with, in the lab. 

Filtering eucalyptus dyes. Mucky business.

Filtering eucalyptus dyes in the lab.

I realized while doing this work that labware is so perfect for working with fibre. Heat resistant glass beakers and flasks are amazing for cooking fibre in liquid, and a magnetic stirrer and hot plate does most of the work. Also, they are marked with measurements - very convenient. The highest setting on the hot plate brings liquids to a perfect, gentle, low boil and maintains it, so you can walk away and leave it. I'm really surprised more textile dye labs are not set up with scientific labware, they're such well designed utilitarian objects, which look cool, too. Easy to clean, stain resistant and durable (pretty smash-proof). I know what I'll have in my studio when I set up a permanent one again, post-grad school.

So, the disaster that happened last night was a cause of great anxiety, and a whole lot of work today. It was a result of working in a shared lab space: someone (I don't know who) turned the warm water bath up to 70˚C and left it like that. The warm water bath is supposed to be set at body temperature (37˚C) for warming up cell culture media to use in your culture dishes, to feed your cells. I went into the lab and set up everything I needed to feed my cells at the end of the day yesterday. I put three different flasks of various cell media in the warm water bath, turned it on and left the lab for 20 minutes while the UV light sterilized the flow hood. When I came back, I discovered the warm water bath steaming like a hot tub and all of my media cooking in it, nearly bursting its plastic flasks and popping the lids off them. It was a big WHAT THE FUCK?? moment. Basically, my media was ruined, the proteins in it all denatured (which means cooked and no longer fresh and usable by cells, basically). I had no choice but to feed the denatured media to my cultures, since they were starving, and deal with the fallout today. 

Most of my day today consisted of running around gathering the chemicals to make new bone soup and regular D-10 (the standard cell medium). The majority of my C2C12s died. There are some left, but it'll be a long road to recovery now that they're sitting back in the right medium. I couldn't even bear to look at the marrow cells yet. I'll inspect those later under the digital microscope to be sure of what I'm looking at. Pray with me to Saint Henry, for survivors. Making bone soup completely on my own was nerve-wracking but I managed to do it, with a little help from Stuart. I love that man. Oron was home with his sick daughter and Ionat is out of the country, so I was really on my own with all of this. Once again, I had a chance to empower myself by getting tough work done in the lab using my own wits and whatever resources I could find. So, I have new bone soup, a whole fresh batch. Thankfully I still have one vial of frozen cells to make use of!

Boxes full of cryovials of cells, being brought up from liquid nitrogen.