High-resolution time-lapse imaging of cell response to CDK2-inhibition

One of the major advantages of Nanolive’s label-free imaging is that images can be acquired at high frequency, over long periods of time. This allows us to analyse the responses of cells to drug-perturbation with high temporal precision.

Here, we observe how pre-adipocytes and glioblastoma cells respond to a cyclin-dependent kinase 2 (CDK2) inhibitor. One image was observed every 2 seconds for 20 hours. The resulting images were analysed using Nanolive’s cell metrics tool. The graph on the right shows how the average dry mass of a cell changes throughout the experiment; pre-adipocyte cells are shown in green, and glioblastoma cells are shown in orange.

Pre-adipocyte cells response to CDK2 inhibition

The videos permit several interesting observations. Let us focus on the response of pre-adipocytes first. The initial response to CDK2 inhibition seems to be an increase in the number of protrusions; the cells become more spread across the surface of the dish. This lasts approximately 11 hours. At 11:48 the first instance of cell death occurs. After this point, cells start to detach from the surface, lose dry mass and become rounder. There are clear changes in the nuclear membrane, which appears to thicken. A couple of cells die before the experiment is ended, but many remain alive.

Glioblastoma cells response to CDK2 inhibition

Now, let us compare the response of glioblastoma cells to CDK2 inhibition. The cells are heavily impacted from the start. No cell protrusions are visible and apoptotic bodies are already present in some cells. Cells start to die 6 hours after the inhibitor was added. Death occurs during mitosis, which makes sense as CDK2 is an important checkpoint in the cell cycle (it drives the progression of cells into the S- and M-phases of the cell cycle). Glioblastoma is a very aggressive type of cancer, which divides rapidly, and this may be one reason why we observe mass cell death, and a sharp decrease in dry mass, around the 7-hour mark. Some cells survive until the end of the experiment, these cells share similar phenotypes to surviving pre-adipocyte cells.

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