In cancer research, it all comes down to a single cell.
Over the past decade, cancer researchers have focused on the fact that an individual cell from a tumor can be used to perform molecular analyzes that reveal important clues about how the cancer developed. , how it spreads and how it can be targeted.
With this in mind, a team of researchers from Brown University has developed an advanced method for isolating single cells from complex tissues. In a study published in Scientific reportsthey show how the approach not only results in high-quality intact single cells, but is also superior to 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 cells from biopsied cancerous tissue to prepare them for analysis, said Anubhav Tripathi, study author and director of biomedical engineering at Brown.
“From a technology perspective, there’s nothing like it on the market right now,” Tripathi said. “This technology will be useful for those looking for answers using genomics, proteomics, transcriptomics. It will not only facilitate these diagnostic and therapeutic investigations, but also save researchers time and effort.”
Tripathi added that beyond clinical applications, the technology will be useful in biomedical applications such as tissue engineering and cell culture.
In single cell analysis, advanced sequencing techniques are used to obtain genetic profiles of individual cells. This particularly applies to cancerous tissues, where rare mutations can lead to metastasis and treatment-responsive outcomes. A major limitation of the clinical translation of single-cell analysis is the difficulty of isolating single cells from complex tissues, said co-author Nikos Tapinos, associate professor of neurosurgery and neuroscience at Brown.
Tapinos described a typical workflow using the example of a brain tumor: a piece of the tumor would be removed in an operating room and taken to a lab. There, researchers would use a process involving enzymes to extract nucleic acids from bulk tissue samples and then perform bulk genetic sequencing.
This process results in low-resolution and potentially inaccurate gene readouts and poor detection of rare cell types, Tapinos said. The consequences of losing this information can be profound, he noted, including the possibility of misdiagnosing the patient, creating a significant lag between when the tumor is removed from the patient and when the cells are ready for RNA sequencing.
“There is a huge need for a technology that can remove tissue from the patient and, within minutes, obtain viable, healthy single cells from which RNA can be isolated,” Tapinos said. “That’s exactly what this new process does.”
The advantages of electricity over enzymes
In the new method, a tissue biopsy is placed in a fluid-filled receptacle between two parallel flat electrodes. Instead of enzymes, electric field fluctuations are applied to create opposing forces in the liquid. These forces cause the tissue cells to move in one direction and then in the opposite direction, causing them to separate or cleanly dissociate from each other.
This approach was invented by study author Cel Welch, who holds a fourth-year Brown Ph.D. candidate in biomedical engineering in the laboratory of Tripathi.
“Dr. Tripathi has done a lot of work in his lab using electric fields and microfluidics,” Welch said. “After seeing how electric fields could be used in other diagnostic applications, we came up with the idea of doing something unique with the electric field that had never been done before. Based on the corpus of research on manipulating biological particles, we formulated a hypothesis about how it would work.”
The new process resulted in dissociation of biopsy tissue in as little as 5 minutes, three times faster than leading enzymatic and mechanical techniques described by Tripathi and Welch in a previous study.
The approach also resulted in “good dissociation of tissues into single cells while preserving cell viability, morphology, and cell cycle progression, suggesting utility for sample preparation of tissue samples for single-cell analysis. direct,” the study concluded.
According to the researchers, the new approach is, at a minimum, 300% more efficient than even the most optimized techniques using simultaneous chemical and mechanical dissociation.
Another benefit of the process, Welch said, is the compactness of the device they created: “In the traditional workflow, you have to use several different lab instruments, like a centrifugation instrument, which cost several thousand dollars each. This approach to single-cell sample preparation requires only one device.”
The research team has filed a US and worldwide patent application for the device and associated methodology, with assistance from 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-scale tissue biopsy samples at once, at very low cost.
The new study explained the scientific basis for the process, Welch said. “Now we are working on a new device that is specifically geared towards creating this highly optimized system for using this physical phenomenon.”
“A researcher will be able to simply press a button and, within minutes, have the single cells they need for analysis,” Tapinos said. “It’s really unbelievable.”
Computational approach enables spatial mapping of single-cell data in tissues
E. Celeste Welch et al, Electric field facilitated rapid and efficient tissue dissociation into viable single cells, Scientific reports (2022). DOI: 10.1038/s41598-022-13068-6
Provided by Brown University
Quote: Scientists Develop New Method and Device to Isolate Single Cells Using Electric Fields (July 14, 2022) Retrieved July 14, 2022 from https://phys.org/news/2022-07-scientists- method-device-isolate-cells.html
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