Hu Tu-80 Dilution Series and L-cell Islands

Today at Applied Biophysics I ran an experiment using a new type of cell known as Hu Tu-80. Despite its catchy sounding name, Hu Tu-80 cells are actually a bit sinister- they are human intestinal cancer cells. So far I have worked only with animal cells- from monkeys, dogs, and mice. This new human cell line looked similar to the animal cells I have worked with. Hu Tu-80 cells share some of the same properties as MDCK cells such as their cobblestone-shaped growth, but had more defined cell boundaries similar to that of BSC-1 cells.

This experiment is designed to examine the effects of BSA and Gelatin on varying concentrations of Hu Tu-80 cells. In this experiment, I used a 96W1E+ (which is an electrode array with 96 wells, and one electrode in each well). In the odd columns, I coated the wells with BSA (Bovine Serum Albumin), and the even columns were coated with Gelatin. BSA is used as a protective agent for cells from oxidative damage, and also serves to stabilize components of media such as fatty acids and pyridoxal. Gelatin is made from the collagen of animal parts and is primarily used to improve cell attachment. I let 200 micro liters of these two solutions sit in the wells for about 5 minutes, suctioned out the wells, and then added varying concentrations of Hu Tu-80 cell mixture to each of the wells. The Hu Tu-80 cell solution was diluted with a corresponding amount of regular medium. Here is the layout of my 96 array (the last two columns are empty):

Green columns are BSA coated, yellow columns are Gelatin coated.

L-cell and BSC-1 dilution series results:

Two weeks ago, I performed a dilution series experiment with L-cells and BSC-1 cells. I was testing these two cells types to see if they formed islands like the BSC-1 and MDCK cells did. The result: YES! As the concentration of L-cells increased, L-cell islands became more prevalent. Dr. Keese stained a row of my array to show how the number of islands increased. In the following pictures, you can see these stained wells:    ( *L-cells are the dark, circular cells* )

Pure BSC-1

Light inoculation - .0625 concentration of L cells

L-cells islands become more prevalent at .25 concentration

L-cell islands become as populous as BSC-1 cells

Pure L-cells

Next week, I will be working with Dr. Keese to begin setting up the time lapse equipment for our next experiment! We will be using the time lapse video to watch the formation of these islands in real time. Can't wait!!