Tissue culture technniques routinely utilise substances that are directly or indirectly non-human animal byproducts, like the matrigel mimesis of the extracellular matrix. Martigel is derived from an Engelbreth-Holm-Swarm (EHS) mouse sarcoma (tumour). Primary cultures of kertinocytes are grown on a feeder layer of 3T3 cells, which are a mouse fibroblast cell line. They quite literally feed by growing the extracellular matrix - or vital environment the keratinocytes can proliferate in. The 3T3s are treated so that they don’t proliferate.

These cross species environments and processes make radical crossings from a cultural perspective and defy cultural practices of clear seperateness and containment, undermining received taxonomies and orders of things.

Alan (Xiaodong Zhuang) works in Roy Bicknell’s lab and has been of enourmous help in showing me how things work and explaining proctocols and proceedure to me. He’s been doing all the tissue culture work so far or supervising my work. On Monday he set up a culture of HUVEC (Human Umbilical Vein Endothelia Cells) to grow on a matrigel so we could observe their growth into tubules. Matrigel is a substance that mimics the extracellular matrix (ECM) that mamalian cells require to sustain and supprt them, it’s their contextual environment.

Alan trypsonised the Human Umbilical Vein Endothelia Cells at 37 degrees - the body temperature necessary for the enzymatic actions of typson to engage, dissassociating them from the dish and eachother. Media with foetal calf serum was added to halt the tryson action and then spun in the centrifuge so that the pellet of cells could be resuspended in new media without the tryson. Then the cells were counted so that the correct numbers of cells could be suspended and seeded onto the matrigel. I followed the protocol until the maths bit - which I got but doubted I could of done so assuredly if I’d done the protocol alone.  The cells were then placed into the incubator.

For some reason I didn’t get the bit of infomration that expalined th time scale of the cells tubule formation and came back too late. The cells had already been at their optimum and were now in various stages of cell death, so i missed the boat on that one. It’s tricky been such a non-scientist in this context, it’s impossible for others to have a decent grasp of my knowledge base and I probably nod my head too much like I know what is going on.  Plus the business and goodwill of others makes me not want to keep anyone way from their proper job any more then they need to be, so I’ve been slow on the upatake a few times. However there might be an opportunity to run it again and to make a time lapse movie of the cell actions and organisation.

This week I resumed the residency following an extened trip in the USA, a mixture of retreat and holiday.

I have begun two cell culture activites:

• In Prof. Roy Bicknell’s lab in the Medical School I’m learning to culture endothelial cells - SEND cells - a murine skin endothelium cell line. Endothelial cells line the interior of blood vessels, some endothelial cultures self organise into tube like structures when grown in a matrix gel - which is why I am interested in them.

• In the The School of Dentistry biomaterials I’m culturing fibroblasts - 3T3 cells - another murine cell line, towards seeding domestic spiders webs with them to see if they function as a viable scaffold for cell growth.

It’s quite something going from lab to lab, trying to pick up the varying practices and proceedures around working with cell cultures and asceptic tecniques - and nervewracking as a non scientist. The principles are the same but the interpretations of them and behaviours around practices of containment, sterlility and exclusion very different.

some wear gloves, some don’t

some wear lab coats, some don’t

some douse everything in ethanol reagardless of the type of work involved, some distinuguish between cell line, primary, human, DNA and so on.

some listen to music, some don’t

some say lids of TC flasks etc. face down when you place them on a surface, some say lids face up

So I am intrigued as I cross labs and learn techniques, behaviours, practises and superstitions.