https://scholars.lib.ntu.edu.tw/handle/123456789/628854
標題: | A Systemic Approach to Isolate, Retrieve, and Characterize Trophoblasts from the Maternal Circulation Using a Centrifugal Microfluidic Disc and a Multiple Single-Cell Retrieval Strategy | 作者: | SHIN-YU LIN Lu, Li-Kuo Hsu, Wei-Fan Peng, Wei-Chieh Tseng, Hua-Wei Li, Chih-Chi Chen, Chen-Lin Huang, Guan-Syuan CHIEN-NAN LEE ANDREW WO |
關鍵字: | ENDOTHELIAL-CELLS; BLOOD-CELLS; RARE CELLS; E-CADHERIN; CANCER; EXPRESSION; SIZE | 公開日期: | 14-二月-2023 | 出版社: | AMER CHEMICAL SOC | 卷: | 95 | 期: | 6 | 來源出版物: | Analytical chemistry | 摘要: | Rare cells in the blood often have rich clinical significance. Although their isolation is highly desirable, this goal remains elusive due to their rarity. This paper presents a systemic approach to isolate and characterize trophoblasts from the maternal circulation. A microfluidic rare cell disc assay (RaCDA) was designed to process an extremely large volume of up to 15 mL of blood in 30 min, depleting red blood cells (RBCs) and RBC-bound white blood cells (WBC) while isolating trophoblasts in the collection chip. To minimize cell loss, on-disc labeling of cells with fluorescent immuno-staining identified the trophoblasts. Retrieval of trophoblasts utilized an optimized strategy in which multiple single cells were retrieved within the same micropipette column, with each cell encapsulated in a fluid volume (50 nL) separated by an air pocket (10 nL). Further, whole-genome amplification (WGA) amplified contents from a few retrieved cells, followed by quality control (QC) on the success of WGA via housekeeping genes. For definitive confirmation of trophoblasts, short-tandem repeat (STR) of the WGA-amplified content was compared against STR from maternal WBC and amniocytes from amniocentesis. Results showed a mean recovery rate (capture efficiency) of 91.0% for spiked cells with a WBC depletion rate of 99.91%. The retrieval efficiency of single target cells of 100% was achieved for up to four single cells retrieved per micropipette column. Comparison of STR signatures revealed that the RaCDA can retrieve trophoblasts from the maternal circulation. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/628854 | ISSN: | 0003-2700 | DOI: | 10.1021/acs.analchem.2c04260 |
顯示於: | 應用力學研究所 |
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