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Microbiology JoVE (Open Access) Citable · DOI

Rapid Isolation of Viable Circulating Tumor Cells from Patient Blood Samples

DOI: 10.3791/4248-v
What you'll learn
  • ✓ Prepare halloysite nanotube-coated microtubes for cell capture
  • ✓ Isolate viable circulating tumor cells from patient blood samples
  • ✓ Achieve high-purity CTC capture using E-selectin and epithelial antibodies
  • ✓ Evaluate CTC isolation efficiency and cell viability post-capture
Protocol

Circulating tumor cells are isolated from the blood of cancer patients without inflicting cellular damage. Isolation of tumor cells is accomplished using a bimolecular surface of E-selectin in addition to antibodies against epithelial markers. A nanotube coating specifically promotes cancer cell adhesion resulting in high capture purities.

Difficulty
advanced
Total time
~3-4 hours per sample (tube preparation ~2 hours; cell isolation ~1-2 hours)
Biosafety
BSL-2

Steps

1
Prepare halloysite nanotube solution

Prepare a halloysite nanotube suspension for coating microtubes. This solution serves as the adhesive substrate for cancer cell capture.

▶ 01:15
2
Coat microtube inner surface with nanotubes

Apply halloysite nanotube solution to the inner surface of microtubes to establish a biocompatible coating that promotes selective cancer cell adhesion.

▶ 01:48
3
Prepare microtubes for cell isolation

Functionalize nanotube-coated microtubes with E-selectin and epithelial marker antibodies to enable specific circulating tumor cell capture.

▶ 03:10
4
Prepare blood samples for cell isolation

Process patient blood samples to remove contaminants and prepare them for loading into prepared microtubes.

▶ 04:05
5
Isolate circulating tumor cells from blood

Incubate blood samples in functionalized microtubes to allow selective adhesion and capture of viable circulating tumor cells.

▶ 05:07
6
Evaluate CTC isolation efficiency and purity

Assess the quantity, viability, and purity of isolated circulating tumor cells using microscopy and validation assays.

▶ 06:18
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