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Imprinted NanoVelcro Microchips for Isolation and Characterization of Circulating Fetal Trophoblasts: Toward Noninvasive Prenatal Diagnostics.
Hou, Shuang; Chen, Jie-Fu; Song, Min; Zhu, Yazhen; Jan, Yu Jen; Chen, Szu Hao; Weng, Tzu-Hua; Ling, Dean-An; Chen, Shang-Fu; Ro, Tracy; Liang, An-Jou; Lee, Tom; Jin, Helen; Li, Man; Liu, Lian; Hsiao, Yu-Sheng; Chen, Peilin; Yu, Hsiao-Hua; Tsai, Ming-Song; Pisarska, Margareta D; Chen, Angela; Chen, Li-Ching; Tseng, Hsian-Rong.
Affiliation
  • Hou S; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Chen JF; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Song M; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Zhu Y; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Jan YJ; Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine , Guangzhou, China.
  • Chen SH; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Weng TH; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Ling DA; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Chen SF; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Ro T; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Liang AJ; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Lee T; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Jin H; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
  • Li M; PacGenomics , Agoura Hills, California 91301, United States.
  • Liu L; PacGenomics , Agoura Hills, California 91301, United States.
  • Hsiao YS; PacGenomics , Agoura Hills, California 91301, United States.
  • Pisarska MD; Department of Obstetrics and Gynecology, Cathay General Hospital , Taipei, Taiwan 106.
  • Chen A; Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center , Los Angeles, California 90048, United States.
  • Chen LC; Department of Obstetrics and Gynecology, University of California, Los Angeles , Los Angeles, California 90095, United States.
  • Tseng HR; California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles , Los Angeles, California 90095-1770, United States.
ACS Nano ; 11(8): 8167-8177, 2017 08 22.
Article in En | MEDLINE | ID: mdl-28721719
ABSTRACT
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.
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Full text: 1 Database: MEDLINE Main subject: DNA / Comparative Genomic Hybridization Type of study: Diagnostic_studies / Guideline Limits: Adolescent / Adult / Female / Humans / Male Language: En Year: 2017 Type: Article
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Full text: 1 Database: MEDLINE Main subject: DNA / Comparative Genomic Hybridization Type of study: Diagnostic_studies / Guideline Limits: Adolescent / Adult / Female / Humans / Male Language: En Year: 2017 Type: Article