Scientists have developed a new method and device for isolating individual cells using an electric field

In order for cancer researchers to analyze and sequence individual cells from complex tissues such as tumors, these cells must first be separated from bulk samples. By: Sel Welch

In cancer research, everything comes down to a single cell.

Over the past decade, cancer researchers have turned their attention to the fact that a single tumor cell can be used for molecular analysis that provides important clues about how the cancer developed, how it spreads, and how it can be targeted.

With this in mind, a team of researchers at Brown University has developed an advanced way to isolate individual cells from complex tissues. In a study published in Scientific reportsthey show how this approach not only results in high-quality, intact single cells, but also outperforms standard isolation methods in terms of labor, cost, and efficiency.

The challenge was to develop technology that would allow researchers to more quickly and easily isolate cancer cells from biopsies fabrics to prepare it for analysis, said Anubhav Tripathi, study author and director of Brown’s Department of Biomedical Engineering.

“From a technology standpoint, there’s nothing like it on the market right now,” Tripathi said. “This technology will be useful for those looking for answers through genomics, proteomics, transcriptomics — it will not only facilitate these diagnostic and therapeutic studies, but also save researchers time and effort.”

Tripathi added that apart from clinical applications, the technology will be useful in biomedical applications such as tissue engineering and cell cultures.

U unicellular analysis, advanced sequencing techniques are used to obtain genetic profiles of individual cells. This is especially true in cancerous tissues, where rare mutations can cause metastases and treatment outcomes. A major limitation in the clinical translation of single-cell analysis is the difficulty of isolating single cells from complex tissues, said co-author Nikos Tapinas, associate professor of neurosurgery and neurology at Brown.

Tapinas described a typical workflow using the example of a brain tumor: a piece of the tumor is removed in the operating room and taken to the laboratory. There, researchers will use a process involving enzymes to extract nucleic acids from bulk tissue samples and then perform bulk genetic sequencing.

This process results in potentially inaccurate, low-resolution genetic readings and poor detection of rare cell types, Tapinas said. The consequences of losing this information can be profound, he noted, including the possibility of misdiagnosing a patient, creating a significant lag between the time a patient’s tumor is removed and the cells ready for RNA sequencing.

“There is a huge need for a technology that allows us to remove tissue from a patient and, in a matter of minutes, result in viable, healthy individual cells from which RNA can be extracted,” Tapinas said. “That’s exactly what this new process does.”

Advantages of electricity over enzymes

In the new process, a tissue biopsy is placed in a fluid-filled container between two parallel plate electrodes. Instead of enzymes, electric field fluctuations are used to create opposing forces in the liquid. These forces cause tissue cells to move in one direction and then in the opposite direction, causing them to separate cleanly or separate from each other.

This approach was invented by study author Sal Welch, a fourth-year Brown Ph.D. candidate in biomedical engineering in Tripathi’s lab.

“Dr. Tripathi has done a lot of work in his lab using electric fields and microfluidics,” Welch said. “After we saw how electric fields could be used in other diagnostic applications, we had the idea to do something unique with an electric field that had never been done before. Building on existing research on the manipulation of biological particles, we formulated a hypothesis about how it will work.”

The new process resulted in biopsy tissue being dissociated in just 5 minutes – three times faster than the leading enzymatic and mechanical methods described by Tripathi and Welch in a previous study.

The approach also resulted in “good tissue dissociation into single cells while preserving cell viability, morphology and cell cycle progression, suggesting utility for sample preparation of tissue samples for direct single-cell analysis,” the study concluded.

According to the researchers, the new approach is at least 300% more efficient than even the most optimized methods using simultaneous chemical and mechanical dissociation.

Welch said that another advantage of the process is the compactness of the device they created: “In a traditional workflow, you have to use several different laboratory instruments, such as a centrifuge, each of which costs several thousand dollars. This approach to preparing a sample from a single cell requires only one device.”

The research team has applied for a US and global patent for the device and related methodology with the help of Brown Technology Innovations, the university’s technology transfer office.

The samples used in the study were bovine liver tissue, triple negative breast cancer cells, and human clinical glioblastoma tissue. The research team is currently refining the technology and developing a device that will be able to quickly and efficiently process several different types of small tissue biopsy samples simultaneously at a very low cost.

The new study sheds light on the scientific basis of the process, Welch said. “We are now working on a new device that is specifically focused on creating this highly optimized system to exploit this physical phenomenon.”

“A researcher will be able to simply press a button and get a single in a few minutes cells they need an analysis,” Tapinos said. “It’s really amazing.”

The computational approach enables spatial mapping of single-cell data in tissues

Additional information:
E. Celeste Welch et al., Electric Field Facilitated Rapid and Efficient Dissociation of Tissues into Viable Single Cells, Scientific reports (2022). DOI: 10.1038/s41598-022-13068-6

Citation: Scientists develop new method and device to isolate individual cells using electric fields (2022, July 14) Retrieved July 14, 2022, from isolate-cells.html

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