When a tumor is present in the human body, cells naturally eliminate it during the formation and growth stages. Circulating tumor cells(CTCs) are the name given to these remaining fragments in the bloodstream (CTCs).
CTCs are highly valued in medical research because they can alert doctors to the presence of a tumor before imaging. Working backward, scientists can use CTCs to determine the approximate size and location of a primary mass in order to begin treatment as soon as possible.
The most significant limitation of CTC isolation is its scarcity. As a result, numerous techniques have been developed and are constantly being improved to improve the efficacy of circulating tumor cell separation and enumeration. They have demonstrated the ability to not only detect the presence of a tumor but also to provide us with its core information.
CTC Separation
The process of isolating CTCs from residual blood cells is known as CTC separation. Red blood cells (RBCs), white blood cells (WBCs), and other substances can all impair the clarity of subsequent experimental results.
CTCs must be isolated and purified before they can be studied properly. CTC Enrichment refers to the process of isolating and purifying CTCs.
Uses For CTC Separation
The detection of tumor cells at various stages of circulation through the body is the primary research field for CTC separation. CTC separation aids oncologists in early cancer diagnosis because the cells can be detected before any imaging shows signs of a tumor.
CTC separation can also assist doctors in determining what type of treatment a patient should receive for their cancer; this targeting method makes treatment more individualized and effective.
Other advantages include the ability to research how cancer spreads and metastasizes, how various cancers affect the body, and how to determine the threat level of a tumor from the bloodstream. This means that CTC separation has a high potential to improve patient treatment and outcomes.
There are several approaches to eliminating contamination and ensuring the most enriched sample possible.
CTC Process
The most common method for CTC separation is to use antibodies to target the surface antigens of tumor cells. Major histocompatibility complex (MHC) molecules in the immune system can identify unknown cells and bind peptide chains to them, allowing other cells to delete them. This process can be artificially replicated to distinguish CTCs from other cells.
This can be difficult because CTCs exhibit varying levels of epithelial markers and different antigens can form on similar cells. As a result, they are more difficult to retrieve or mark with antibodies. They also risk being destroyed by the immune system before being extracted.
When attempting to separate by size or density, CTC uses blood filtration and Density Gradient Centrifugation. Filtration has a high separation efficiency and is easily automated, but it can become inaccurate due to cell accumulation and unwanted cells squeezing through filters. Density gradient centrifugation aids in the grouping of CTCs but leaves them heavily contaminated by RBCs.
Limitations
CTCs are not completely saturated in blood samples. These cells are extremely difficult to isolate due to their scarcity. Many current methods struggle to purify the sample without the use of additional steps. Even after cleaning with a density gradient and microfluidics, a sample can still contain more than 90% RBCs.
When sorting, these RBCs take up space that would normally be filled by more CTCs; they also make it difficult to culture rare cells due to the physical space they take up. The presence of these cells contaminates enriched CTC samples, making it more difficult to secure results for downstream applications.
A New Approach To Colon Carcinoma Circulating Tumor Cell (CTC) Detection Using Pluribead
We present a novel method for screening colon carcinoma circulating tumor cells (CTCs) with pluriBead containing a tumor-associated EpCAM antibody.
This method is based on the technology of non-magnetic cell separation. It does not necessitate any sample preparation. EpCAM-pluriBead® can be directly injected into a whole blood sample. The method is also suitable for isolating single cells from various biological fluids. Its sensitivity can also be increased by increasing the sample volume.
Following molecular-genetic experiments to determine their mutation status, the bound EpCAM-positive colon carcinoma cells can be easily included. In the case of colon carcinoma, K-ras mutation status is a tool for predicting response to anticancer therapy. As a result, the new method can be regarded as a quick and effective tool for early cancer detection.
Steps Involved In Tumor Study
- Labeling
Add EpCAM-pluriBead® to your sample
- Mixing
30 minutes of gentle incubation (recommended with pluriPlix®).
- Isolation
Captured target cells separation is done via appropriate sieves.
- Washing
Use wash buffer to clean sieve. Lyse cells with Trizol®.
- CTC Processing
Approaches for the study of cancer cells:
– RNA/DNA isolation
– Cell culture experiments
Conclusions And Perspectives
Using EpCAM-pluriBead®, we developed a new method for detecting circulating colon tumor cells in the blood. As a result, the captured cells are suitable for additional molecular-genetic screening for specific markers associated with tumor formation.
The developed “in situ immunoblastspcr” method eliminates the need for preliminary RNA/DNA isolation and can significantly reduce analysis time.Finally, the goal of this work is to improve the sensitivity of the method by increasing the sample volume, varying the cell tumor lines, and/or changing the antibody specificity.
Check Out Pluriselect’s Cell Separation Technology Today
We have your needs covered for cell separation and isolation. If you are searching for a way to improve the quality and viability of sorted cells while dramatically saving time and reducing costs, look no further than Pluribead.
CTCs have the potential to help patients throughout their cancer journey, from diagnosis to treatment selection, post-treatment/surgery monitoring, and follow-up.
Despite the fact that a large amount of research has been accelerated in the field of these disseminated tumor cells, their scarcity has limited research. We anticipate the development of cell separation and enrichment fusion techniques that will aid in the prevention of cell loss.
A variety of specific markers is also likely to improve enrichment results, as cells that can avoid EpCAM selection may also be captured. Given their enormous potential to help change the enigmatic situation of solid tumors, we can conclude that CTCs will undoubtedly become an unavoidable part of solid tumor malignancies in the near future.
Please contact us if you have any questions, and try our products as soon as possible.
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