Institute of Microelectronics, Peking University, Beijing 100871, China.
Lab Chip. 2019 Mar 13;19(6):974-983. doi: 10.1039/c8lc01335g.
In melanoma surgery, it is difficult to identify residual scattered tumor cells at the surgical margin because of invasive growth. Mohs surgery, widely applied to increase the cure rate and decrease the recurrence rate of melanoma, involves examination of the tissue for tumor cells after tissue removal. Here, we established a liquid biopsy platform for rapid (<5 h), sensitive examination of residual tumor cells at the margin after Mohs surgery using clinical samples from patients with pigment nevus for a demonstration. The design involved highly sensitive, selective rare target cell separation from surgical margin lavage fluid (SMLF) through micropore-arrayed filtration. High recovery rates (86.7% ± 16.3% and 72.7% ± 46.7%, respectively) for separation of spiked 5 A375s (cultured human melanoma cells) and 1 A375 from 1 mL PBS were achieved for this platform. Detection of SMLF samples from patients with pigment nevus was performed, and many (66-7420) Melan-A-positive target cells were successfully recovered and identified, demonstrating the application performance of this rapid liquid biopsy for Mohs surgery in clinical practice. Moreover, a high-selectivity separation of larger target A375 cells from smaller background Jurkat cells was achieved with a high enrichment factor (4.2 ± 1.1). In clinical practice, high selectivity contributes to effective depletion of red blood cells (RBCs), thus ensuring verification of target cells from samples with severe RBC contamination. Furthermore, target cells were obtained with high purity (2.7-35.2%). The capability of this method for rare-cell separation with a high recovery rate and good selectivity may facilitate improvement of performance of Mohs surgery for real clinical practice, including shortening examination time and increasing detection sensitivity.
在黑色素瘤手术中，由于浸润性生长，很难在手术切缘处识别残留的散在肿瘤细胞。Mohs 手术广泛应用于提高黑色素瘤的治愈率和降低复发率，涉及在组织切除后检查组织中的肿瘤细胞。在这里，我们建立了一种液体活检平台，用于快速（<5 小时）、灵敏地检查 Mohs 手术后手术切缘冲洗液（SMLF）中残留的肿瘤细胞，并用患者的色素痣临床样本进行了验证。该设计涉及通过微孔阵列过滤，从手术切缘冲洗液中高度敏感、选择性地分离稀有靶细胞。该平台实现了对 1 mL PBS 中分别掺入的 5 个 A375s（培养的人黑色素瘤细胞）和 1 个 A375 的高回收率（分别为 86.7%±16.3%和 72.7%±46.7%）。对色素痣患者的 SMLF 样本进行了检测，并成功回收和鉴定了许多（66-7420）个 Melan-A 阳性靶细胞，证明了这种快速液体活检在临床上应用于 Mohs 手术的效果。此外，该平台还实现了从较小的背景 Jurkat 细胞中高选择性地分离较大的靶细胞 A375，具有较高的富集因子（4.2±1.1）。在临床实践中，高选择性有助于有效去除红细胞（RBC），从而确保从 RBC 严重污染的样本中验证靶细胞。此外，还获得了高纯度（2.7-35.2%）的靶细胞。该方法具有高回收率和良好选择性的稀有细胞分离能力，可能有助于提高 Mohs 手术的性能，包括缩短检查时间和提高检测灵敏度。
