Isolating Single Gene Cells

Using flow cytometry to separate cells from the microenvironment guides treatment decisions

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If there were any doubt remaining about the outlook for single cell gene expression, it vanished last fall with the National Institute for Health's (NIH's) announcement of a $90 million support of single cell gene studies and genomic sequencing.

On the forefront of this movement is NanoString Technologies, Inc., a provider of life science tools for translational research and molecular diagnostic products, and BD Biosciences, a segment of BD, a global medical technology company, today announced a collaboration agreement for the development of a single cell isolation and analysis workflow.

Under the agreement, the companies will jointly develop a workflow using the NanoString nCounter Analysis System (including the nCounter Single Cell Assay) and the BD Flow Cytometry cell sorter product line (emphasizing the new BD FACSJazz Cell Sorting System). The combined workflow will enable single cell gene expression analysis for research applications such as oncology, immunology and stem cell research. Collaboration activities will also include the development of materials documenting the workflow protocol, as well as co-hosting meetings and webinars to educate scientists about the single cell workflow.

Though oncology and stem cell research comprise the bulk of BD Biosciences and NanoString Technologies work right now, application possibilities are virtually endless and range from tumor biopsies to marine ocean samples. Regardless of end use, the long-term goal is to accelerate the move towards personalizing health to the cellular level by understanding the link between cell variation, tissue and organ function, and emergence of disease.

"Maximizing both the quantity and quality of data that can be extracted from a single cell is critical to the emerging field of single cell biology. The nCounter Analysis System can analyze entire gene pathways and provides a highly precise and reproducible digital output, making it ideally suited to the task," said Brad Gray, president and chief executive officer, NanoString Technologies. "The nCounter Analysis System and the BD FACSJazz Cell Sorting System can together provide a powerful and efficient workflow for single cell gene expression analysis."

"Our collaboration with NanoString Technologies furthers BD's commitment to providing researchers advanced solutions for cell analysis and isolation," said Alberto Mas, President, BD Biosciences. "We believe this new sorting workflow will complement the recent and very rapid advances in genomic studies that value the requirement for greater sample integrity for critical single cell analysis."

Ultimately, the technology holds the potential to predict clinical outcomes and rely less on invasive surgery, said Clark Mason, PhD, senior director of global planning at BD Bioscience.

"The tumor microenvironment has a direct affect on how tumors evolved in the body," he explained. "Metastasis is what kills patients most commonly. Cell interaction in the microenvironment correlates with the microenvironment for the primary tumor. This technology offers a finer dissection of the actual tumor and the amount of material you need lessens with the more sensitive techniques. If you use less of the tumor, you don't need to rely on such invasive surgical techniques. It's cheaper and much less stressful for the patient."

Flow Cytometry Advantage
According to Mason, the use of flow cytometry makes their product unique.

"You can't get more sensitive as there's no further definition of life than a single cell," he reasoned. "We used to drop whole liver tissues into a blender for gene expression. Now we're down to complex tissues, like cells of the tumor microenvironment. You can extract it by hand, which is very laborious. You can use lasers, but, by using flow cytometry, you can subject the findings to gene analysis."

The advantages of ensuring the right cell is being analyzed can't be overstated, Mason said.

"When you input into a micromanipulator, you're never sure if another cell or part of the cell is coming along for the ride," he explained. "Flow cytometry isolates the cell, makes sure it's alive and not dying, and then selects the cell for analysis."

NanoString Technologies' nCounter Analysis System is a multi-application digital detection and counting system with a highly automated and simple workflow. The company's single cell gene Expression application provides researchers with a highly flexible and sensitive approach to discovering differences in cell-to-cell gene expression profiles. The application enables up to 800 genes to be detected in a single tube.

"Our software allows users to mine the data more effectively and reveal the trends and deductions that allow them to make the gene expression with specific cells," said Mason. "Once they have the data and see the heat maps, they can look at the particular effect of the cells giving gene certain gene expressions."

The BD FACSJazz Cell  Sorting System is capable of identifying, characterizing and isolating single or multiple cells -- from complex or extremely rare cell populations - and depositing them in 96 and 384 well plates to provide rapid cell isolation, tracking and identification throughout the process.

Robin Hocevar is on staff at ADVANCE. Contact rhocevar@advanceweb.com.


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ibtehal omer,  ms,  nd histopathologyJanuary 03, 2014
sudan, AL


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