A Silicon-based Coral-like Nanostructured Microfluidics to Isolate Rare Cells in Human Circulation: Validation by SK-BR-3 Cancer Cell Line and Its Utility in Circulating Fetal Nucleated Red Blood Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Coral Chip Manufacture
2.3. Cell Spiking Test
2.4. Fetal Nucleated Red Blood Cells (fnRBCs) Capture
2.5. Fluorescence in Situ Hybridization (FISH)
2.6. Captured Cells Recovery
2.7. Whole Genome Amplification (WGA)
2.8. Array Comparative Genomic Hybridization (aCGH)
2.9. Next Generation Sequencing (NGS)
2.10. Short Tandem Repeat (STR) Analysis
3. Results
3.1. Capture Efficiency Estimated by Cell Spiking Test
3.2. Circulating fnRBC Captured by Coral Chip
3.3. FISH
3.4. Captured Cells Recovery
3.5. WGA
3.6. aCGH and NGS Analysis
3.7. STR Analysis
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample No. | No. of Captured SK-BR-3 cells | No. of Falsely Captured Jurkat Cells | Capture Rate for SK-BR-3 Cells (%) | False Capture Rate for Jurkat Cells (%) |
---|---|---|---|---|
1 | 4012 | 7 | 80.24 | 0.0007 |
2 | 4241 | 0 | 84.82 | 0 |
3 | 4683 | 0 | 93.06 | 0 |
4 | 4728 | 1 | 94.56 | 0.0001 |
Mean | 4405 | 2 | 88.17 | 0.0002 |
Case No. | MA (Year) | GA (Week+day) | Pre-acquired Fetal Genetic Condition | cbNIPD | ||
---|---|---|---|---|---|---|
No. of fnRBCs Captured (in 2 mL Maternal Blood) | Non-maternal Genomic Markers Used to Confirm the Fetal Origin of Captured Cells | Validated * Method | ||||
1 | 30 | 27+5 | arr[GRCh37] 9p24.2p23 (2267812_13374304) × 1 dn | 10 | 1. 9p24.2p23 deletion 2. Chr Y | aCGH [pooled 8] |
2 | 38 | 20+6 | arr[GRCh37] 10q25.2q26.12 (114393625_121720948) × 1 dn | 47 | 1. 10q25.2q26.12 deletion 2. Chr Y | aCGH [pooled 13] |
3 | 31 | 25 | arr[GRCh37] 21q22.11q22.3 (35703384_48056450) × 1 dn | 47 | 1. 21q22.11q22.3 deletion 2. Chr Y | aCGH [pooled 15] |
4 | 40 | 18 | arr[GRCh37] 22q11.21 (18894835_21505417) × 1 dn | 18 | 22q11.21 deletion | aCGH [pooled 10] |
5 | 28 | 15+6 | 48,XXY,+18 | 7 | T18 | FISH [4] |
6 | 37 | 13+4 | 47,XY,+18 | 25 | T18 | FISH [10] |
7 | 29 | 16 | 47,XY,+18 | 3 | T18 | FISH [3] |
8 | 34 | 20+6 | 47,XY,+21 | 14 | T21 | FISH [6] |
9 | 43 | 25+6 | 46,XY | 3 | Chr Y | FISH [3] |
10 | 32 | 19 | 46,XY | 2 | Chr Y | FISH [2] |
11 | 29 | 24+6 | 46,XY | 10 | Chr Y | FISH [6] |
12 | 37 | 15 | 46,XY | 10 | Chr Y | FISH [4] |
13 | 28 | 24 | 46,XY | 71 | Chr Y | FISH [22] |
14 | 42 | 24 | 46,XY | 6 | Chr Y | STR analysis [pooled 5] |
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Ma, G.-C.; Lin, W.-H.; Huang, C.-E.; Chang, T.-Y.; Liu, J.-Y.; Yang, Y.-J.; Lee, M.-H.; Wu, W.-J.; Chang, Y.-S.; Chen, M. A Silicon-based Coral-like Nanostructured Microfluidics to Isolate Rare Cells in Human Circulation: Validation by SK-BR-3 Cancer Cell Line and Its Utility in Circulating Fetal Nucleated Red Blood Cells. Micromachines 2019, 10, 132. https://doi.org/10.3390/mi10020132
Ma G-C, Lin W-H, Huang C-E, Chang T-Y, Liu J-Y, Yang Y-J, Lee M-H, Wu W-J, Chang Y-S, Chen M. A Silicon-based Coral-like Nanostructured Microfluidics to Isolate Rare Cells in Human Circulation: Validation by SK-BR-3 Cancer Cell Line and Its Utility in Circulating Fetal Nucleated Red Blood Cells. Micromachines. 2019; 10(2):132. https://doi.org/10.3390/mi10020132
Chicago/Turabian StyleMa, Gwo-Chin, Wen-Hsiang Lin, Chung-Er Huang, Ting-Yu Chang, Jia-Yun Liu, Ya-Jun Yang, Mei-Hui Lee, Wan-Ju Wu, Yun-Shiang Chang, and Ming Chen. 2019. "A Silicon-based Coral-like Nanostructured Microfluidics to Isolate Rare Cells in Human Circulation: Validation by SK-BR-3 Cancer Cell Line and Its Utility in Circulating Fetal Nucleated Red Blood Cells" Micromachines 10, no. 2: 132. https://doi.org/10.3390/mi10020132