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Simpler Lab Cell Production Techniques Devised

Human epithelial cells

Human epithelial cells (Icelandl, Wikimedia Commons)

27 January 2017. A process is being developed that makes it easier to produce healthy and diseased cells in the lab from human specimens for regenerative medicine and research. The techniques designed by pathologists Richard Schlegel and Xuefeng Liu at Georgetown University Medical Center are described in the 26 January issue of the journal Nature Protocols (paid subscription required).

Georgetown University holds a patent on the technology invented by Schlegel and Liu, issued by the U.S. Patent and Trademark Office in March 2016. Schlegel is also a co-founder of the three year-old spin-off company Propagenix in Rockville, Maryland with an exclusive license to the rights to commercialize the technology.

Schlegel, Liu, and colleagues aim to simplify the processes of growing new cells in the lab with samples of cells taken from patients or other human donors. Production of new human cells can be helpful for identifying treatment targets and testing drug candidates for safety or early indicators of efficacy. These techniques are also becoming applicable for regenerating new cells to replace damaged or diseased tissue in patients, which can reduce or eliminate risks of immune-system reactions.

Current cell generation techniques like those using stem cells, say the authors, have important limitations. Cells kept in cultures often have limited life spans, with little if any potential for proliferation. Cell lines kept continuously also can change their genetic characteristics, and thus their biochemical properties. And while induced pluripotent stem cells show promise for growing new cells in the lab, they too can be difficult to maintain, expand, and differentiate into new cells. These limitations are particularly meaningful for precision medicine diagnostics and treatments for cancer.

The researchers call the techniques to address these issues conditional reprogramming, on which they started work in 2011. Condition reprogramming starts with human cells, which can be healthy or cancerous, and are then cultured in feeder cells — irradiated connective tissue cells called fibroblasts — and a compound known as a Rho kinase inhibitor. Rho-kinase proteins regulate a number of cellular functions, including cell division and proliferation, thus blocking their actions encourages cell growth and production. No genetic modifications are needed.

Schlegel, Liu, and colleagues from Georgetown’s Center for Cell Reprogramming already published a number of demonstrations of their processes since 2011. In the new study, the team used conditional reprogramming to produce new epithelial cells, from the lining of hollow organs and glands such as blood vessels and esophagus. These epithelial cells were derived from both fresh and cryopreserved samples kept in super-cooled state. They reported being able to produce some 1 million new cells in 7 days, which could also be continued indefinitely.

Liu says in a university statement that biobanking is a particularly promising application of conditional reprogramming. “We can grow cells, freeze them, thaw them,” notes Liu. “Think about use of such cells for skin replacement, for organ patching, and cancer studies.”

Among the demonstrations of conditional reprogramming is generation of functioning beta cells from the pancreas that produce insulin. People with type 1 diabetes, an autoimmune disorder, cannot produce their own insulin and rely on injections or insulin pumps to maintain their blood glucose levels. “A true cure for this kind of diabetes,” adds Liu, “could be achieved by replacing the lost beta cells with new functional insulin producing cells.”

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