Veterans Affairs

Rotating wall vessel for cell cultures

Inexpensive and less labor intensive approach better suits the needs of the pharmaceutical industry in ADME-Tox screening

Medical & Biotechnology

Scientists at the U.S. Department of Veterans Affairs (VA) have recently developed a suspension culture device and rotation mechanism for the optimal environment for cell studies. The technology is based on zero gravity research and holds promise in providing much-improved results in ADME-Tox screening. The patented technology is available via patent license agreement to companies that would make, use, or sell it commercially.

The three-piece design holds the culture in the middle layer on a breathable surface. Windows in the top piece provide for visual examination and imaging.

To have the useful predictive capability for biologic studies, in vitro cell culture models must reproduce as many features as possible of the in vivo cells or tissues that they are representing. As an example, using screening drugs for hepatoxicity using cultured liver cells is useful to the extent that those cells display the characteristics of cells in the body’s liver. However, most, if not all, differentiated cells derived from tissues lose their specialized features and de-differentiate when grown under traditional petri dish cell culture conditions. Suspension culture, where cells float in the media without adhering to plastic dishes, is the most popular technique for minimizing this problem and maintaining the cell’s specialized features. However, a significant drawback of these approaches is the resulting turbulence and impact against the stirrers damaging the cells. Rotating wall vessels (RWVs) have been used to address these issues.

Hepatocytes grown in an RWV form 3D organoids that maintain their function for at least several weeks. Current RWVs have a large capacity and are well adapted for generation of cell-derived products. But, the vessels can be costly, require expensive electrical hardware to rotate them, are labor-intensive to load, maintain, and harvest, and are poorly amenable to automation. Furthermore, the large numbers of cells and large volume required per vessel can be prohibitively expensive for use in drug screening, where thousands of individual cultures are needed to assay the multiple drugs, each at varying doses and times of exposure.

Addressing the above, VA researchers have developed, a three-piece suspension culture device along with a rotating machine. The hockey puck design of the suspension culture features a middle layer for the culture medium and is made of breathable material. The device holds liquid, cells, biologicals, micro-organisms, and matrices including beads. Multiple ports permit fluid and gas communication in and out of the system.

The roller system keeps the fluid in laminar flow.

While offering a variety of uses, it may be best utilized for screening drugs for hepatoxicity and nephrotoxicity. The RSV device may maintain target cells in a state of differentiation that improves acute toxicity testing and may maintain viable and functional cells for long time periods to allow for evaluation of chronic toxicity. Further, the device may induce pathological states such as shear elevated to a few to several times physiological to mimic pathological flows, such as increased renal tubular flow in many types of chronic kidney disease.

The complete system may be ideal for the study of kidney metabolism, antibody production, cartilage implants, cosmetic components, pancreatic islets, and stem cells.

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