I’m trundling along here in my new lab, still trying to get everything up and running. On the tissue culture front, things have been fraught for some time, what with delays installing the carbon dioxide and nitrogen tanks, with learning how to live in harmony with the experimental prokaryotes sharing the same air space – all while finishing off a few key applications for funding. Finally, however, I thought I had the system working smoothly, and was ready to perform my first real experiment: infecting bladder epithelial cells with patient bacteria isolated from fresh urine.
This is an absolutely straightforward procedure. I’m simplifying it a little bit, but in essence, you seed your cells onto a coated glass surface (such as the chamber slide pictured above), perform the infection and, some time later, wash the cells and then fix and stain them to inspect what the bacteria have done to the cells and vice versa. It will be my bread-and-butter assay, the basis for testing the genetics and cell biology of infection and also trialling some of the interventions we hope to discover and perfect. So in that sense it is also a bottleneck: the basic experiment without which little else can progress.
So I was pretty miffed when it became apparent that Step One of this simple procedure – the step so basic that you wouldn’t normally even spare it a thought – was just not working. In short, I could not get the bloody cells to stick to the slide. Now this isn’t some ultra-finicking primary cell we’re talking about here: it’s ‘T24’, a full-out transformed tumor cell line that’s been growing for yonks, that’s so happy in culture that you have to split it multiple times per week to keep it under control. These are cells that are obviously quite keen to adhere when the notion suits them. They are the Sultans of Stickiness. They have gorgeous, frilly pseudopods that fan out as they probe their environment and inch their way across the plastic surface, forming and dissolving sticky adhesions with ease as they go. I could gaze at them for hours under the microscope, admiring their luscious lamellipodia.
But on glass, the game was completely off. Many cells can’t grow directly on glass, so when that failed, I tried coating with poly-D-lysine, fibronectin, collagen, serum, and attachment factor. All failed: the cells remained round, clinging together desperately in increasingly large clumps that wafted around the slide like seaweed on the ocean floor before dying a tragic death.
The real mystery, however, began when I contacted the lab in Cardiff that had kindly posted me the cells in the first place. Yes, T24 grew just fine on glass, I was told. No, no coating was required. Yes, they were perfectly happy, in fact, on the precise make of slides that I was using.
So, Dear Readers – I throw the question open. What now? Can anyone help solve the mystery? Or will my line of research suffer…a sticky end?