Since my last post, amazing things have happened.
I've been awarded a very generous international artist-in-residence grant from artsnb (the New Brunswick Arts Board). This enabled me to close my crowd funding campaign, because my budget is now at exactly the amount it ideally needed to be for this project. Previous to this, I was coasting on the absolute bare minimum that I needed to be here. What a relief it was to receive word that I was selected for this grant! The New Brunswick Arts Board has been absolutely phenomenal in supporting my art career over the past 13 years, just when I need support the most. We New Brunswick artists are very lucky to have this resource. The things I will be able to do now at SymbioticA will be quite extraordinary.

More good news, too - I have a bone cell! : 



This, my friends, is my very first viable bone cell!
ISN'T IT BEAUTIFUL? It looks sort of like a hummingbird. This is from the rat bone marrow that I extracted from the rat that Henry the cat caught for me. Henry, my hero. The cell is the splatted thing on the left. The ghostly-looking splat stuff is the cell's cytoplasm, which has spread out and adhered itself to the bottom of the pitre dish, meaning that this cell is healthy and growing. The luminous thing on the bottom right with the tentacles is likely another cell as well, but it's unclear how healthy it is because only the nucleus is visible and no cytoplasm. So, it may be dying... or maybe it will grow. I'll know in a few days. If you look back to the splat, you can see a similarly sized nucleus in it, a white roundish thing that is underneath another layer of cell stuff. The long tendrils that reach out into the nothingness are the cell's way of networking with other cells, and when they join, the begin to form a colony, which is what I want. It was a joyous day the day I discovered this cell under the microscope! Just imagine me, like a space explorer, searching the empty universe for life, day after day, to one day stumble across this! No small accomplishment, let me tell you. Primary sourcing cells from recently dead tissue is very, very difficult. My other cell cultures are doing nothing and will just have to be thrown out. I've been struggling to work with this technique because I discovered that ordering cell lines from a supplier is much more expensive than I first imagined: the cell lines themselves are around $500-$700, but with the shipping, it's a total expense of about $2000. $2000 for a bunch of frozen cells in a miniscule vial, shipped in a box kept at -80˚. If it continues to grow and multiply, I will eventually be able to spin down and freeze my own vial of osteoblasts, and save them for later or allow someone else at SymbioticA in the future to use them, contributing to the ongoing research there. When I looked at this cell today with my naked eye through the microscope, it was glowing and radiant. A stunning, inspiring communion with the force of life, through just a single cell.

3D printed scans of Stelarc's arm with the ear implant.

Other exciting things I've been doing include 3D printing miniature looms on Stelarc's amazing, fancy 16micron 3D printer. Stelarc needs no introduction, as far as I'm concerned but just in case: He's the artist who has an ear implanted in his arm, and it's been there since about 2006 or so. He's internationally famous in the BioArt world. He's an amazing, generous man, and is a professor at Curtin University in Perth. He's been teaching me how to use his extremely sophisticated (best in the world) 3D printer so that I can use it to print objects for my bone project.

So, I'm printing looms for weaving. Each loom is approximately 3cm x 3cm, and I've printed some miniature loom tools as well, like shuttles and needles. The looms so far are different models of simple frame and peg looms, which will allow me to weave basic cloths. I will get more elaborate with printing and assembling looms after I experiment with these ones, perhaps even print a full floor loom model in miniature and assemble it. Why the miniature looms?

My newest research has led me to discover that silk fibre is not only compatible with human biological systems (meaning the body and cells like it, it will break down in the body over time and it's even been used successfully in bone grafts already), but it also carries "indicators" that communicate with bone cells about how to differentiate, meaning it basically has programming for bone cells on each silk filament. What this means is that you don't have to use a special differentiation medium, like bone soup, to grow osteoblasts. The silk will tell them what to do, so you can grow osteoblasts in regular old cell medium. If this isn't the most radical and incredible thing ever, I don't know what is. Mind = blown. I ordered unspun silk filament thread and will weave some into miniature cloths and then will insert the entire loom with the cloth still on it into a pitre dish and culture cells on it. Imagine how amazing this will be! I'm also going to weave some of my decellularized intestine on it, as well as surgical sutures (which, by the way, are either made out of SILK or GUT). I have ample sutures waiting to be used, and will begin prepping my silk yarns tomorrow. I pick up my printed looms from Stelarc on Friday and then start weaving. I also printed some already woven structures, like a woven ball and a basket weave, to try them out in culture dishes as well, to see what happens. I'm not sure if cells will grow on that kind of polymer or not, because it's not organic (so probably not). Ultimately, I'm hoping not, because I want my cells to just grow in the shape of the cloths I weave, not the looms. I will post pics of my miniature looms once I have them. Here are some of the things that Stelarc has printed as experiments for himself:

A 3D printed, functioning miniature bike chain. He gave me this as a gift.

A 3D printed, functioning miniature wrench.

I was also amazed and delighted to discover that preparing silk for a cell culture is exactly the same as prepping it for dyeing: washing in boiling water with soda ash for 1 hour, 2x. The only difference is that I'll dry the silk in a fume hood that's turned on overnight and autoclave it (cook it in extremely high temperature steam) to kill absolutely everything that might possibly contaminate it. Tonight, I am making my own soda ash (or sodium carbonate). I discovered that you can do this by simply putting baking soda (sodium bicarbonate) in the oven at 200˚C for 1 hour. Ta-dah, you have soda ash. It's just fantastic how much of my tissue engineering work is a combination of science, kitchen witchery and textile know-how. This particular aspect of things thrills me.

My eucalyptus dye, before (left) and after (right) adding mordant.

Some of my bone slides being stained.

Speaking of dyeing, I've been playing with my eucalyptus bark dye, prepared as an histological stain for tissue sections on glass slides. I also dyed a shirt with it. It stained the tissue on the microscopic slides exactly the same as the cotton shirt, but interestingly enough, I didn't have to use a mordant on the shirt to get it to hold the dye, whereas I did have to add a mordant in the lab to get it to properly dye the bone and other tissue sections on the slides. Bone seems particularly hard to dye. There are plenty of mordants in the lab, as well as oxidizers which help catalyze the dye. I used sodium iodate to oxidize the eucalyptus, as well as iron as a mordant. It worked well. However, the dye liquor was not really as saturated as it could have been and so everything I used it on turned a light coffee colour. I'd like it darker, so I'll boil down the liquor more and saturate the dye/stain.

This bloody gash is eucalyptus sap.

I've also learned more about the mixing and uses of histological stains, such as haematoxylin, which is a natural stain derived from the heart of haematoxylin wood. Haematoxylin stains the nuclei of cells black. Other stains (both natural and chemical) will stain different types of tissue, such as collagen. So, I've experimented with a number of these and now have a beautiful box of glass slides of tissues that I've sliced, dyed and mounted myself. They are treasures. Again, I will post more photos of those later when I have some. What I discovered about the eucalyptus dye was that it didn't discriminate between tissue types - it just dyed everything a coffee brown. I was hoping to make some great eucalyptus lab discovery, but alas...

Macro shot of eucalyptus sap, which looks remarkably like the potassium ferricyanide crystals I was growing, here: http://ossificatorium.wordpress.com/photo-gallery/

OK, here are my C2C12s now, differentiated into multiple nuclei cells (versus cells with one nuclei) - you can see them form longer, stretched out cells now. You can also see the cytoplasm around some of them. I told my peeps at SymbioticA today that I'm about as good at feeding and maintaining cells (which need to be fed every two days) as I am at watering plants and looking after animals, meaning kind of not really. These guys just got fed after five days. I'll try to do better!

Mighty Mouse cells!

And, my entire assortment of tissue cultures in the incubator at present - that's a lot of hungry cells: