5 years ago

Imprinted NanoVelcro Microchips for Isolation and Characterization of Circulating Fetal Trophoblasts: toward Noninvasive Prenatal Diagnostics

Imprinted NanoVelcro Microchips for Isolation and Characterization of Circulating Fetal Trophoblasts: toward Noninvasive Prenatal Diagnostics
Li-Ching Chen, Margareta D. Pisarska, Shang-Fu Chen, Yu Jen Jan, Hsiao-Hua Yu, An-Jou Liang, Shuang Hou, Ming-Song Tsai, Yazhen Zhu, Lian Liu, Tom Lee, Tzu-Hua Weng, Hsian-Rong Tseng, Min Song, Tracy Ro, Angela Chen, Peilin Chen, Szu Hao Chen, Man Li, Yu-Sheng Hsiao, Dean-An Ling, Jie-Fu Chen, Helen Jin
Circulating fetal nucleated cells (CFNCs) in maternal blood offer an ideal source of fetal genomic DNA for noninvasive prenatal diagnostics (NIPD). We developed a class of nanoVelcro microchips to effectively enrich a subcategory of CFNCs, i.e., circulating trophoblasts (cTBs) from maternal blood, which can then be isolated with single-cell resolution by a laser capture microdissection (LCM) technique for downstream genetic testing. We first established a nanoimprinting fabrication process to prepare the LCM-compatible nanoVelcro substrates. Using an optimized cTB-capture condition and an immunocytochemistry protocol, we were able to identify and isolate single cTBs (Hoechst+/CK7+/HLA-G+/CD45–, 20 μm > sizes > 12 μm) on the imprinted nanoVelcro microchips. Three cTBs were polled to ensure reproducible whole genome amplification on the cTB-derived DNA, paving the way for cTB-based array comparative genomic hybridization (aCGH) and short tandem repeats analysis. Using maternal blood samples collected from expectant mothers carrying a single fetus, the cTB-derived aCGH data were able to detect fetal genders and chromosomal aberrations, which had been confirmed by standard clinical practice. Our results support the use of nanoVelcro microchips for cTB-based noninvasive prenatal genetic testing, which holds potential for further development toward future NIPD solution.

Publisher URL: http://dx.doi.org/10.1021/acsnano.7b03073

DOI: 10.1021/acsnano.7b03073

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