Detection of circulating tumor cells from cryopreserved human sarcoma peripheral blood mononuclear cells

Abstract

Circulating tumor cells (CTCs) enter the vasculature or lymphatic system after shedding from the primary tumor. CTCs may serve as “seed” cells for tumor metastasis. The utility of CTCs in clinical applications for sarcoma is not fully investigated, partly owing to the necessity for fresh blood samples and the lack of a CTC-specific antibody. To overcome these drawbacks, we developed a technique for sarcoma CTCs capture and detection using cryopreserved peripheral blood mononuclear cells (PBMCs) and our proprietary cell-surface vimentin (CSV) antibody 84-1, which is specific to tumor cells. This technique was validated by sarcoma cell spiking assay, matched CTCs comparison between fresh and cryopreserved PBMCs, and independent tumor markers in multiple types of sarcoma patient blood samples. The reproducibility was maximized when cryopreserved PBMCs were prepared from fresh blood samples within 2 hours of the blood draw. In summary, as far as we are aware, ours is the first report to capture and detect CTCs from cryopreserved PBMCs. Further validation in other types of tumor may help boost the feasibility and utility of CTC-based diagnosis in a centralized laboratory.

Isolation of circulating tumor cells in a preclinical model of osteosarcoma: Effect of chemotherapy

“…Using a method based on isolation by size of tumor cells (ISET), Chinen et al [29] detected CTCs in sarcoma patients. More recently, Li et al [30] confirmed the detection of CTCs in cryopreserved human peripheral blood mononuclear cells and two studies analysing CTCs by FISH in small cohorts of osteosarcoma patients revealed a potential positive association between CTC count, disease progression and poor prognosis [31] , [32] . The present report is the first study investigating the regulation of CTC release after chemotherapy.…”

Establishment of a Model of Microencapsulated SGC7901 Human Gastric Carcinoma Cells Cocultured with Tumor-Associated Macrophages

“…Blood samples (15 mL) were collected from healthy donors. Peripheral blood mononuclear cells (PBMCs) were isolated according to a previous publication [ 13 ]. PBMCs were plated in three 6-well plates (1 × 10 6 cells/well) and incubated for 3 h in RPMI1640 (Gibco) supplemented with 1% (v/v) penicillin-streptomycin and 10% FBS in an incubator at 37°C and 5% CO 2 at 100% relative humidity.…”

Soft‐tissue sarcoma in adults: An update on the current state of histiotype‐specific management in an era of personalized medicine

“…In the future, postoperative surveillance strategies should be tailored to the specific histiotype given the differences in tumor biology and patterns of recurrence. The value of molecular surveillance with circulating tumor cells is also being investigated 257,258 …”