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Abstract
BackgroundCirculating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes.MethodsFour cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain.ResultsOur results suggest that primary/immortalized cell lines stored in SBTS remain ~90 % viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS.ConclusionsIn this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs after days of storage. Therefore, we suggest an effective and economical transportation of cancer patient blood samples containing live CTCs can be achieved.
Highlights
Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology
While further studies on the potential use of the sugar based cell transportation solution (SBTS) to preserve live CTCs in transporting blood samples is needed, we suggest that cells can be kept viable when stored long term in blood at ambient temperatures
A recent addition to the arsenal of clinical blood testing procedures in oncology practices is the enumeration of circulating tumor cells (CTCs)
Summary
Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. The primary mechanism of metastatic spread begins with dissemination, or shedding, of cancerous epithelial cells from tumor sites into the circulation. These circulating tumor cells (CTCs) travel throughout the body, adhere to organ vascular beds, infiltrate the tissue, grow and impair organ function [1,2,3]. Despite ~106 tumor cells are being shed into the circulation per gram of tumor tissue every 24 h, less than 1 % of shed CTCs remain alive 24 h after dissemination [8,9,10,11] This loss of CTC viability has been attributed to many factors including fragility, shear stresses in the circulation, anoikis and Stefansson et al BMC Cancer (2016) 16:300 destruction by the immune system [12,13,14,15,16]. Studies imply that the injected cancer cells that form primary tumors have different biological properties than their progeny populating the metastatic tumors [17, 18]
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