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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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The heart, an organ critical to survival, gets nourishment not from the blood it pumps but from a separate circulation system known as coronary circulation. This is the shortest circulation in the body and is responsible for supplying the heart with the nutrients it needs to function effectively.
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Early diagnosis and treatment can often cure cancer. However, even with treatment, residual cells called cancer stem cells (CSC) might remain, often causing tumor recurrence. These cancer stem cells possess the potential for self-renewal and multi-lineage differentiation and are often responsible for the therapeutic resistance displayed in most cancers.
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In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells
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Sensitive and easy screening for circulating tumor cells by flow cytometry.

Alexia Lopresti1, Fabrice Malergue2, François Bertucci1,3

  • 1Predictive Oncology Laboratory, Cancer Research Center of Marseille (CRCM), Inserm U1068, CNRS UMR7258, Institut Paoli-Calmettes, Aix Marseille Université, Marseille, France.

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Detecting circulating tumor cells (CTCs) is challenging. This study presents a rapid, one-hour flow cytometry protocol for sensitive CTC detection from small blood volumes, aiding cancer diagnosis.

Keywords:
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Area of Science:

  • Oncology
  • Biotechnology
  • Cell Biology

Background:

  • Circulating Tumor Cells (CTCs) offer valuable insights into solid tumors but are difficult to detect due to rarity and lack of specific biomarkers.
  • Current flow cytometry (FC) methods for CTC detection are often lengthy, requiring pre-enrichment and extended staining, hindering clinical efficiency.
  • The heterogeneity of CTCs, including epithelial-mesenchymal plasticity, complicates reliable detection.

Purpose of the Study:

  • To develop a rapid and efficient flow cytometry protocol for sensitive Circulating Tumor Cell (CTC) detection.
  • To overcome the limitations of time-consuming pre-enrichment and staining procedures in existing FC-based CTC assays.
  • To establish a method for identifying CTCs from small blood volumes that is independent of their plasticity.

Main Methods:

  • Simultaneous fixation, permeabilization, and staining of cells to reduce processing time.
  • Utilized low-speed flow cytometry acquisition with cell size and pan-cytokeratin expression as discriminators to eliminate the need for pre-enrichment.
  • Applied the protocol to blood samples from donors with and without known malignant diseases.

Main Results:

  • The developed protocol significantly reduced the CTC detection procedure to under one hour with minimal manipulation.
  • The method demonstrated high recovery of target cells, irrespective of their epithelial-mesenchymal plasticity.
  • The protocol successfully predicted cancer patient samples, indicating its potential as a diagnostic tool.

Conclusions:

  • This proof-of-concept study establishes a sensitive and rapid flow cytometry tool for CTC detection from small blood volumes.
  • The streamlined protocol enhances the efficiency of CTC analysis, paving the way for broader clinical application.
  • This approach provides a foundation for further in-depth analyses of CTCs for cancer management.