Three-Dimensional Scaffold Chip with Thermosensitive Coating for Capture and Reversible Release of Individual and Cluster of Circulating Tumor Cells

Abstract

Tumor metastasis is attributed to circulating tumor cells (CTC) or CTC clusters. Many strategies have hitherto been designed to isolate CTCs, but there are few methods that can capture and gently release CTC clusters as efficient as single CTCs. Herein, we developed a three-dimensional (3D) scaffold chip with thermosensitive coating for high-efficiency capture and release of individual and cluster CTCs. The 3D scaffold chip successfully combines the specific recognition and physically obstructed effect of 3D scaffold structure to significantly improve cell clusters capture efficiency. Thermosensitive gelatin hydrogel uniformly coated on the scaffold dissolves at 37 °C quickly, and the captured cells are gently released from chip with high viability. Notably, this platform was applied to isolate CTCs from cancer patients' blood samples. This allows global DNA and RNA methylation analysis of collected single CTC and CTC clusters, indicating the great potential of this platform in cancer diagnosis and downstream analysis at the molecular level.

Beyond Capture: Circulating Tumor Cell Release and Single‐Cell Analysis

“…CTC purity can be enhanced to 88–98% via two rounds with capture/release, and the improved purity allowed for subsequent cell expansion and mutational analysis on epidermal growth factor receptor (EGFR) gene. Gelatin is another potential thermoresponsive material for CTC release, which undergoes the gel‐to‐sol transition beyond its upper critical solution temperature around 33 °C . Chen and co‐workers developed a 3D scaffold chip with gelatin coating for capture and gentle release of individual and cluster CTCs .…”

“…Gelatin is another potential thermoresponsive material for CTC release, which undergoes the gel‐to‐sol transition beyond its upper critical solution temperature around 33 °C . Chen and co‐workers developed a 3D scaffold chip with gelatin coating for capture and gentle release of individual and cluster CTCs . Gelatin hydrogel was uniformly coated on the scaffold, allowing for the modification of anti‐EpCAM Ab to capture CTCs.…”

“…b) Gelatin coated‐3D scaffold chip for capture and release of individual and cluster CTCs. Reproduced with permission . Copyright 2017, American Chemical Society.…”

Circulating tumor microemboli: Progress in molecular understanding and enrichment technologies

“…To facilitate the release of captured cells/clusters, the same group of researchers have recently introduced gelatin hydrogel coated 3D scaffold chip. Gelatin hydrogel is thermosensitive and melts at physiological temperature (37ᴼC) thus enables easy retrieval of viable CTCs/CTM that are suitable for a range of downstream molecular analyses (Cheng et al, 2017).…”

“…Anti-EpCAM functionalized 3D macroporous PDMS scaffold chip High efficiency capture by combining two strategies: chaotic cell migration through spatially distributed macroporous structure, and enhanced binding with antibodies due to nanorough scaffold surfaces Flow rate: 100 µl/min (Cheng et al, 2016) Gelatin hydrogel coating of chip Melting of gelatin hydrogel at 37 ᵒC enables easy retrieval of viable CTCs/CTM (Cheng et al, 2017)…”