Isolation of circulating fetal trophoblasts by a four-stage inertial microfluidic device for single-cell analysis and noninvasive prenatal testing

Huang，Yifang1; Yu，Sheng2; Chao，Shuzhe3; Wu，Limei4; Tao，Maliang5; Situ，Bo5; Ye，Xinyi5; Zhang，Ye5; Luo，Shihua5; Chen，Weishan5; Jiang，Xiujuan5; Guan，Guofeng3; Zheng，Lei6

2020-11-24

10.1039/d0lc00895h

LAB ON A CHIP   影响因子和分区

1473-0197

1473-0189

Noninvasive detection of circulating fetal cells carrying the entire fetal genome is a promising way for prenatal testing of genetic diseases. However, ideal approaches for efficient separation of these valuable cells are not available. Here, a novel inertial microfluidic chip (CelutriateChip 1) is developed for ultra-fast, label-free enrichment of circulating trophoblasts (CTBs) from the whole blood samples of pregnant women. The unique structural design of the four-stage curved channel in CelutriateChip 1 enables CTBs with larger size to be efficiently separated from the blood samples under the effect of inertial and Dean drag forces. The transition of the target cells among the stages enables CelutriateChip 1 to achieve one or two orders of magnitude higher throughput compared to single channel inertial microfluidic chips. After optimization of conditions, CTBs can be recovered from 2 mL of whole blood within 5 min with an average recovery efficiency ranging from 52.3% to 65.8% and high white blood cell depletion (99.95%). CTBs collected from the chip can be isolated at the single-cell level and used for downstream immunofluorescence staining and genetic genotyping. Clinical tests are performed on 30 pregnant women and the results demonstrate that CTBs are obtainable in 86.67% of pregnancy cases. A single-base variant in the HBB gene can be accurately detected by sequencing of rare CTBs. This simple, antibody-free and low-cost approach holds promise for obtaining rare CTBs for prenatal detection of various genetic diseases.

SCI ; EI

英语

其他

National Natural Science Foundation of China[81871735]

Biochemistry & Molecular Biology ; Chemistry ; Science & Technology - Other Topics ; Instruments & Instrumentation

Biochemical Research Methods ; Chemistry, Multidisciplinary ; Chemistry, Analytical ; Nanoscience & Nanotechnology ; Instruments & Instrumentation

WOS:000592314900001

ROYAL SOC CHEMISTRY

20204909582592

Chemical detection ; Drag ; Microfluidic chips ; Molecular biology ; Microfluidics ; Blood

Structural Design, General:408.1 ; Biological Materials and Tissue Engineering:461.2 ; Biology:461.9 ; Microfluidics:632.5.1 ; Chemistry:801

CHEMISTRY

2-s2.0-85096884855

Scopus

1.Department of Laboratory Medicine and, Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China. nfyyzhenglei@smu.edu.cn and Department of Clinical Laboratory, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi Zhuang Autonomous Region, China
2.Department of Laboratory Medicine and, Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China. nfyyzhenglei@smu.edu.cn and Department of Clinical Laboratory, The Third People's Hospital of Shenzhen, Southern University of Science and Technology, National Clinical Research Center for Infectious Diseases, Shenzhen, 518100, China
3.Shenzhen Genflow Technologies Co.,Ltd.,Shenzhen,518000,China
4.Department of Clinical Laboratory,Guangzhou Twelfth People's Hospital,Guangzhou,510620,China
5.Department of Laboratory Medicine and,Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors,Nanfang Hospital,Southern Medical University,Guangdong Province,Guangzhou,510515,China
6.Department of Laboratory Medicine and,Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors,Nanfang Hospital,Southern Medical University,Guangdong Province,Guangzhou,510515,China

Huang，Yifang,Yu，Sheng,Chao，Shuzhe,et al. Isolation of circulating fetal trophoblasts by a four-stage inertial microfluidic device for single-cell analysis and noninvasive prenatal testing[J]. LAB ON A CHIP,2020,20(23):4342-4348.

Huang，Yifang.,Yu，Sheng.,Chao，Shuzhe.,Wu，Limei.,Tao，Maliang.,...&Zheng，Lei.(2020).Isolation of circulating fetal trophoblasts by a four-stage inertial microfluidic device for single-cell analysis and noninvasive prenatal testing.LAB ON A CHIP,20(23),4342-4348.

Huang，Yifang,et al."Isolation of circulating fetal trophoblasts by a four-stage inertial microfluidic device for single-cell analysis and noninvasive prenatal testing".LAB ON A CHIP 20.23(2020):4342-4348.