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Cell Isolation

Cell isolation is a highly significant step in cancer research as well as related investigations because it allows researchers to observe tumor biology, including its microenvironment. Findings from these studies enable scientists to develop drugs and other treatment strategies against specific forms of cancer.

The purpose of cell isolation is to collect specific and viable cells from solid tumors, healthy and diseased tissues in a manner that maintains the biological phenotypes and functions of these cells.

The widely used techniques for cell isolation include enzymatic digestion, mechanical dissociation, gradient centrifugation, magnetic-activated cell sorting (MACS), fluorescence-activated cell sorting (FACS), and single-cell isolation techniques such as laser-capture microdissection and microfluidic devices.

Cell isolation is a significant step in cancer research...

Enzymatic Digestion

Enzymatic Digestion

Enzymatic digestion is a commonly employed technique to dissociate tumor tissues into single-cell suspensions. Enzymes that are typically used include collagenase, trypsin, hyaluronidase, and DNase I. These enzymes function to degrade the extracellular matrix (ECM) so that individual cells can be collected.

The process starts with mincing of the tumor tissue into smaller pieces, which are then incubated at 37°C in a solution with any of the aforementioned enzymes. To ensure that the tissue bits are exposed uniformly to the enzymes, gentle agitation of the mixture is performed. After enzymatic digestion, the undigested debris are removed through filtering with a cell strainers. The sample is then washed to remove residual enzymes, followed by centrifugation to obtain the cell pellet.

This method is advantageous in that high cell yield and preservation of cellular phenotypes can be assured. However, the methodology needs careful optimization to prevent over-digestion which can affect cell surface markers or inflict other cellular damages.

Mechanical Dissociation

Mechanical Dissociation

This technique involves physically disrupting tumor tissues to break them down into smaller fragments or cell suspensions.

Tumor tissues are macerated using a mortar and pestle or mechanically sheared using syringes and needles. Other mechanical devices can also be used to slice or blend tissues into smaller pieces. There are also automated apparatuses designed to accomplish mechanical dissociation in a standardized procedure.

This method is advantageous when dealing with tissues resistant to enzymatic digestion. It is also beneficial since potential enzyme-related alterations are avoided.

Gradient Centrifugation

Gradient Centrifugation

Gradient centrifugation is a technique that uses Ficoll or Percoll to separate cells based on their density. It is commonly used for the enrichment of cancer cells, immune cells and other cell populations after collection from a heterogeneous tumor sample.

After mechanical or enzymatic tissue dissociation, the cell suspension is layered onto the density gradient medium and then centrifuged. This step enables the separation of cells based on their density, allowing the collection of specific cells from distinct layers.

Magnetic-Activated Cell Sorting (MACS)

Magnetic-Activated Cell Sorting (MACS)

MACS uses magnetic beads that are coated with antibodies targeting surface markers expressed on the desired cells. The dissociated cell suspension is incubated with magnetic beads conjugated to antibodies. Target cells bind to the beads and are retained in a magnetic field, while non-target cells are washed away. The magnetic field is then removed to collect the purified cells.

Fluorescence-Activated Cell Sorting (FACS)

Fluorescence-Activated Cell Sorting (FACS)

FACS is a laser-based method that allows precise isolation of specific cell populations based on their fluorescence-labeled surface markers. The dissociated cell suspension is stained with fluorescently tagged antibodies targeting specific markers. A flow cytometer sorts the cells by analyzing their fluorescence intensity and light scattering properties.

Laser-Capture Microdissection (LCM)

Laser-Capture Microdissection (LCM)

LCM is a highly specialized method for isolating cells directly from tumor tissue sections using a laser microscope. Thin tissue sections are prepared and placed on slides. A laser beam selectively cuts and lifts cells of interest from the surrounding tissue, which are then collected for downstream analysis.

Microfluidic-Based Isolation

Microfluidic-Based Isolation

Microfluidic devices use small-scale fluid dynamics to isolate single cells based on size, deformability, or specific surface markers. Tumor tissues are dissociated, and the cell suspension is passed through a microfluidic chip. Channels or filters in the device separate cells based on physical or biochemical properties.

Immunoaffinity-Based Isolation

Immunoaffinity-Based Isolation

This technique involves the use of antibodies specific to cell surface markers to capture target cells. Tumor cell suspensions are incubated with antibodies immobilized on plates, beads, or columns. Target cells bind to the antibodies, while non-target cells are washed away.

Sources: Piwocka et al. (2024), Duan et al. (2013).

Matched Samples

PBMCs

PBMCs (peripheral blood mononuclear cells) | Click here to read more about PBMCs.

PBMCs

PBMCs (peripheral blood mononuclear cells) | Click here to read more about PBMCs.

Biofluids

Biofluids (sputum, saliva, aspirate, swabs, urine, stool, synovial fluid, cerebrospinal fluid) | Click here to learn about our Animal Clinical Samples

Biofluids

Biofluids (sputum, saliva, aspirate, swabs, urine, stool, synovial fluid, cerebrospinal fluid) | Click here to learn about our Animal Clinical Samples

Normal Adjacent Tissue

Normal Adjacent Tissue

Lymph nodes

Lymph nodes

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More on request

Available Services for
Characterization

FACS (fluorescence-activated cell sorting)

qPCR (quantitative Polymerase Chain Reaction)

RNAseq (RNA sequencing)

Sterility

MHC (Major Histocompatibility Complex) typing

More on request

We are About Speed & Quality

Cells are isolated from sourced samples on demand.

Fresh samples are processed at our facilities and cryopreserved cells are shipped on dry ice or liquid nitrogen worldwide.

We typically ship cells that are in stock within two days.

We ship cells Europe-wide within 24-48 hours, and can ship intercontinentally with dry shipper.

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To place an order or inquire about our services, please contact our sales team through the contact form on our website or via email by clicking Contact Us below. We’ll work with you to better understand your specific needs and provide a customized solution.

References

Duan, J. J., Qiu, W., Xu, S. L., Wang, B., Ye, X. Z., Ping, Y. F., Zhang, X., Bian, X. W., & Yu, S. C. (2013). Strategies for isolating and enriching cancer stem cells: well begun is half done. Stem cells and development22(16), 2221–2239. https://doi.org/10.1089/scd.2012.0613

Piwocka, O., Musielak, M., Ampuła, K. et al. (2024). Navigating challenges: optimising methods for primary cell culture isolation. Cancer Cell Int 24 (28). https://doi.org/10.1186/s12935-023-03190-4