Biophysical isolation and identification of circulating tumor cells.
Biophysical isolation and identification of circulating tumor cells.
复制标题
DOI：
10.1039/c7lc00038c
复制
发表时间：
2017-04-11
期刊：
影响因子：
6.1
通讯作者：
Di Carlo D
中科院分区：
文献类型：
--
作者：
Che J;Yu V;Garon EB;Goldman JW;Di Carlo D
从血液中分离和计数循环肿瘤细胞（CTC）对于确定患者预后和监测治疗是重要的。基于对细胞表面标志物的亲和力的方法已经应用于纯化（通过免疫分离）和鉴定（通过免疫荧光）CTC。然而，与肿瘤异质性和演变相关的细胞生物标志物表达的可变性以及抗体探针的交叉反应性长期以来使CTC富集和免疫染色复杂化。在这里，我们报告了一个真正的无标记的高通量微流控方法来分离，计数，并在一个集成的微流控设备上的生物物理特性的CTC的特征。涡旋介导的变形性细胞计数法（VDC）由富集大CTC的初始涡旋区域组成，随后释放到使细胞变形的下游流体动力学拉伸区域中。用高速相机对细胞变形进行可视化和定量显示了来自16个IV期肺癌样品的大（直径>15 μm）和可变形（纵横比>1.2）CTC群体，与污染血细胞和从健康患者分离的罕见大细胞相比，其通过增加的变形性而清楚地区分。VDC技术证明，相对于标准免疫荧光（71.4%），高于健康基线（93.8%）的推定CTC的阳性检出率相当。自动化允许在样品采集后<1小时内从10 mL小瓶血液中完全计数CTC，而标准方法需要4小时以上。此外，细胞被释放到任何收集容器中用于进一步的下游分析。VDC显示出在没有标记的情况下精确CTC计数的潜力，并证实了血液中循环的大CTC的独特的高度可变形的生物物理特性。集成的涡旋富集-变形性细胞计数装置可以通过机械表型分析自动计数血液中的循环肿瘤细胞
Isolation and enumeration of circulating tumor cells (CTCs) from blood is important for determining patient prognosis and monitoring treatment. Methods based on affinity to cell surface markers have been applied to both purify (via immunoseparation) and identify (via immunofluorescence) CTCs. However, variability of cell biomarker expression associated with tumor heterogeneity and evolution and cross-reactivity of antibody probes have long complicated CTC enrichment and immunostaining. Here we report a truly label-free high-throughput microfluidic approach to isolate, enumerate, and characterize the biophysical properties of CTCs on an integrated microfluidic device. Vortex-mediated Deformability Cytometry (VDC) consists of an initial vortex region which enriches for large CTCs, followed by release into a downstream hydrodynamic stretching region which deforms the cells. Visualization and quantification of cell deformation with a high-speed camera revealed populations of large (>15 μm diameter) and deformable (aspect ratio >1.2) CTCs from 16 stage IV lung cancer samples, that are clearly distinguished by increased deformability compared to contaminating blood cells and rare large cells isolated from healthy patients. VDC technology demonstrated a comparable positive detection rate of putative CTCs above healthy baseline (93.8%) with respect to standard immunofluorescence (71.4%). Automation allows full enumeration of CTCs from a 10mL vial of blood within <1 hr after sample acquisition, compared with 4+ hours with standard approaches. Moreover, cells are released into any collection vessel for further downstream analysis. VDC shows potential for accurate CTC enumeration without labels and confirms the unique highly deformable biophysical properties of large CTCs circulating in blood. The integrated Vortex enrichment – Deformability Cytometry device may enable automated enumeration of circulating tumor cells from blood by mechanophenotyping
登录
查看更多内容
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
9.7
作者：
Darling EM;Di Carlo D
通讯作者：
Di Carlo D
影响因子：
5.7
作者：
HARDINGHAM, JE;KOTASEK, D;DOBROVIC, A
通讯作者：
DOBROVIC, A
DOI：
10.1002/pros.22625
发表时间：
2013-06
期刊：
The Prostate
影响因子：
--
作者：
Chen CL;Mahalingam D;Osmulski P;Jadhav RR;Wang CM;Leach RJ;Chang TC;Weitman SD;Kumar AP;Sun L;Gaczynska ME;Thompson IM;Huang TH
通讯作者：
Huang TH
DOI：
10.1007/s00249-013-0888-y
发表时间：
2013-05-01
影响因子：
2
作者：
Kiessling, T. R.;Herrera, M.;Losert, W.
通讯作者：
Losert, W.