Cell Line Development & Engineering

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Cell Line Development

The development of cell lines following a stable transfection step for the purpose of gene expression or recombinant protein production is an important method used by researchers. Generating cell lines has often been inefficient and affected by inaccurate limited dilution methods or the shear stress caused by conventional flow cytometer and traditional high-pressure cell sorters. NanoCellect Biomedical provides cell sorting solutions that allow us to sort cells and establish stable cell lines using gentle low-pressure microfluidics.

This protects cell viability after sorting and improves sorting efficiency by increasing the number of viable clones growing per plate. Contaminate and biohazard–free, the WOLF cell sorter’s rapidly exchangeable cartridge and tubing set eliminates carryover between samples and allows for quick and easy cleanup. More than a cell printer, the WOLF is a FACS (fluorescence-activated cell sorting) instrument capable of sorting based on 5 optical parameters that provide well-by-well index cellular data.

WOLF Benefits

Developing cell lines requires to sort sensitive cells. The ability to sort cells in the lab when they are ready ensures an efficient process to improve cell viability and clonal outgrowth.

The simplest way to ensure no cross-contamination between cell lines is to replace the entire system fluidics at once with a new disposable cartridge. No instrument clean-up required.

Compared to traditional droplet-based cell sorters running under high-pressure, the WOLF provides an alternative sorting method where cells do not undergo a decompression shock before being deposited in the well.

Even for flow cytometry beginners, the WOLF has an intuitive and basic software workflow. It takes a day of training before novice users are up and running in their own lab.

Many laboratories do not have space for a large, conventional cell sorter. Save on space by implementing the WOLF into a cell line development workflow, at only 2 cubic feet. The compact design fits in a biosafety cabinet for sterile cell sorting.

The WOLF instrument setup is rapid and automated. When followed by a gentle sort and no lengthy instrument clean up, the overall process is simple and straightforward.

N1 Single-Cell Dispenser

The N1 Single-Cell Dispenser is an add-on accessory for the WOLF^® cell sorter used for single-cell deposition of viable cells into 96- or 384- well plates. Typically, wells are pre-filled with cell culture media, while the same media is used as the sheath buffer to ensure cell viability. Each well can receive anywhere from 1 to 100 cells and the whole plate can be processed as fast as 9 min and 33 min for 96- or 384- well plates, respectively. The pairing of the low-pressure WOLF^® cell sorter and the N1 single-cell dispenser greatly improves cell integrity from that of conventional sorters, providing higher populations of viable and healthy clones, therefore facilitating stable cell line generation.

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Recombinant protein production in CHO Cells

To produce therapeutic and recombinant proteins, the most widely used mammalian expression system in industrial production is gene amplification using dihydrofolate reductase-deficient (DHFR−) CHO cells (Chinese hamster ovary mammalian cells) with DHFR-mediated gene amplification. Following standard molecular biology plasmid construction methods and stable transfection of cells (such as the host cell line CHO-DG44), the expression of a recombinant protein become stable. The next step requires sorting single-cells to establish recombinant protein producing clones. With traditional cytometers, the number of CHO cell colonies is low due to cell death with high-pressure cell sorting systems. With low-pressure cell sorting, the WOLF results in up to 95% single cell deposition efficiency and more clonal outgrowth.

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Sorting Stem Cell Cultures

Induced pluripotent stem cells and embryonic stem cells have traditionally been very difficult to sort as they are very fragile. Conventional cell sorters cannot be efficiently used to isolate these cells as only a fraction of them survive high-pressure cell sorting steps. Additionally, the stress induced during this process results in stem cell differentiation which is undesirable when working and maintaining stem cell lines. The microfluidic-based WOLF Cell Sorter exhibit higher cell survival after sorting and increased clonal outgrowth. CRISPR-modified hiPS cells selected with WOLF® Cell Sorter exhibit increased viability and genomic integrity.

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Antibody Discovery

Antibodies now represent a growing class of therapeutic molecules for immuno-oncology and immunotherapies. The process of discovering antibodies relies on immunization, isolation of B cells from blood but also the identification of specific antibody producing cells. Typically, antibodies are cloned and express in a variety of host cells followed by the single-cell plating into 96-w and 384- well plates. The WOLF cell sorter gently sorts high antibody titer producing cells. For more information, visit of Antibody Discovery Application page.

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The WOLF is a technology that enables small companies to do things they could not do before.” “The microfluidics capability is something that I have been excited about. The nature of how the WOLF works is being extremely attractive to us. Some of the experiments we’d like to do in the future could not be done on any other device.

Justin Chartron, Ph.D. Protabit LLC

Neurons are extremely delicate so the selection step should be the least disturbing of all if we want to have reliable results. When we tried the WOLF, we were extremely surprised that our cells were very healthy and viability was above 90%, sometimes above 95% which is very impressive.

Dorota Skowronska-Krawczyk, M.Sc.,Ph.D. University of California Irvine

A great machine that has helped us take our cell-engineering pipeline to the next level!

Ronan O'Connell Rice University

The NanoCellect team and technical specialist have been awesome. After two test runs with very involved training/assistance, we have been very pleased with the results. The equipment is intuitive and the sorts have been very clean.

B. Davenport, Ph.D. University of Colorado

NanoCellect Customer Support team has been fantastic in helping us get up and running. The instrument is a helpful tool in our cell line development workflow.

Douglas Austen Takeda Pharmaceuticals

The NanoCellect customer support team is really helpful, knowledgeable and efficient; the WOLF is a great tool in our stem cell research workflow

Helen Miranda, Ph.D. Case Western Reserve University