Conditional cell immortalization using ROCK-inhibitor

by Alexey Bersenev on January 15, 2012 · 0 comments

in methods

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Maintaining the primary cells in culture sometimes is very difficult task. Some normal and malignant cells types just don’t grow well in culture. It makes very difficult cell-based diagnostics and therapy development. Recently, the group of researchers proposed the method, which allows to overcome this problem. They called it “conditional immortalization”.

From Science Daily:

“We tried breast cells and they grew well. We tried prostate cells and their growth was fantastic, which is amazing because it is normally impossible to grow these cells in the lab,” Schlegel says. “We found the same thing with lung and colon cells that have always been difficult to grow.”

“In short, we discovered we can grow normal and tumor cells from the same patient forever, and nobody has been able to do that,” he says. “Normal cell cultures for most organ systems can’t be established in the lab, so it wasn’t possible previously to compare normal and tumor cells directly.”

Using Rho kinase (ROCK) inhibitor and feeder cell layer, Richard Schlegel’s group was able to propagate primary normal and malignant cells for long time. Basically, they created “stem cell-like” conditions. Interestingly, these conditions didn’t push selection for adult- or cancer stem cells from the tested tissues and cells didn’t undergo transformation:

…in contrast to the selection of rare stem-like cells, the described growth conditions can generate 2 × 106 cells in 5 to 6 days from needle biopsies, and can generate cultures from cryopreserved tissue and from fewer than four viable cells. Continued cell proliferation is dependent on both feeder cells and Y-27632, and the conditionally reprogrammed cells (CRCs) retain a normal karyotype and remain nontumorigenic.

This sounds very exciting! I think, more research should be done to confirm the absence of transformation events in immortalized cells.

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