Highly sensitive and rapid isolation of fetal nucleated red blood cells with microbead-based selective sedimentation for non-invasive prenatal diagnostics. (23rd August 2018)

• Record Type: Journal Article
• Authors: Wei, Xiaoyun; Ao, Zheng; Cheng, Lin; He, Zhaobo; Huang, Qinqin; Cai, Bo; Rao, Lang; Meng, Qianfang; Wang, Zixiang; Sun, Yue; Liu, Wei; Zhang, Yuanzhen; Guo, Shishang; Guo, Feng; Zhao, Xing-Zhong
• Is Part Of: Nanotechnology. Volume 29:Number 43(2018)
• Publication Date: 2018-08-23
• Subjects: fetal nucleated red blood cells -- non-invasive prenatal diagnosis -- microbead-based centrifugal separation -- cell release; Nanotechnology -- Periodicals; Nanotechnology -- Periodicals; Nanotechnology; Publications périodiques; Nanotechnologies; Periodicals; 620.5
• DOI: 10.1088/1361-6528/aad8c4
• Languages: English
• ISSNs: 0957-4484

Abstract

Abstract: Non-invasive prenatal diagnostics (NIPD) has been an emerging field for prenatal diagnosis research. Carrying the whole genome coding of the fetus, fetal nucleated red blood cells (FNRBCs) have been pursued as a surrogate biomarker traveling around in maternal blood. Here, by combining a unique microbead-based centrifugal separation and enzymatic release, we demonstrated a novel method for FNRBC isolation from the blood samples. First, the gelatin-coated silica microbeads were modified with FNRBC-specific antibody (anti-CD147) to capture the target cells in the blood samples. Then, the density difference between microbead-bound FNRBCs and normal blood cells enables the purification of FNRBCs via an improved high-density percoll-based separation. The non-invasive release of FNRBCs can then be achieved by enzymatically degrading the gelatin film on the surface of the microbeads, allowing a gentle release of the captured target cells with as high as 84% efficiency and ∼80% purity. We further applied it to isolate fetal cells from maternal peripheral blood. The released cells were analyzed by real-time polymerase chain reaction to verify their fetal origin and fluorescent in situ hybridization to detect fetal chromosome disorders. This straightforward and reliable alternative platform for FNRBC detection may have the potential for realizing facile NIPD.