Cell Isolation as a significant step in preclinical research
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.
Cell isolation is a significant step in cancer research...
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.
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 Extra-Cellular 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
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.
Considerations When Obtaining Animal Clinical Samples
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.
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.
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.
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).
Made-to-order Dissociated Tumor Cells
Dogs
Unit Size: 1 million cells/vial
# | CATALOG NR. | TUMOR TYPE |
|---|---|---|
1 | DTC1001 | Lymphoma |
2 | DTC1002 | Mast Cell Tumor |
3 | DTC1004 | Melanoma |
4 | DTC1005 | Hemangiosarcoma |
5 | DTC1006 | Hepatocellular Carcinoma |
6 | DTC1007 | Bladder Cancer |
7 | DTC1009 | Ear Tumor |
8 | DTC10010 | Thyroid Carcinoma |
9 | DTC10011 | Nasal Tumors |
10 | DTC10012 | Oral Melanoma |
11 | DTC10013 | Soft Tissue Sarcoma |
12 | DTC10014 | Eye Tumor |
13 | DTC10016 | Bone Cancer |
14 | DTC10017 | Kidney Cancer |
15 | DTC10018 | Adrenal Gland Tumor |
16 | DTC10019 | Skin Cancer |
17 | DTC10020 | Mammary Carcinoma |
Cats
Unit Size: 1 million cells/vial
# | CATALOG NR. | TUMOR TYPE |
|---|---|---|
1 | DTC1001 | Lymphoma |
2 | DTC1002 | Mast Cell Tumor |
3 | DTC1004 | Melanoma |
4 | DTC1005 | Hemangiosarcoma |
5 | DTC1006 | Hepatocellular Carcinoma |
6 | DTC1007 | Bladder Cancer |
7 | DTC1009 | Ear Tumor |
8 | DTC10010 | Thyroid Carcinoma |
9 | DTC10011 | Nasal Tumors |
10 | DTC10012 | Oral Melanoma |
11 | DTC10013 | Soft Tissue Sarcoma |
12 | DTC10014 | Eye Tumor |
13 | DTC10016 | Bone Cancer |
14 | DTC10017 | Kidney Cancer |
15 | DTC10018 | Adrenal Gland Tumor |
16 | DTC10019 | Skin Cancer |
17 | DTC10020 | Mammary Carcinoma |
Matched Samples
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
Normal Adjacent Tissue
Lymph nodes
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
