High-throughput isolation of fetal nucleated red blood cells by multifunctional microsphere-assisted inertial microfluid
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High-throughput isolation of fetal nucleated red blood cells by multifunctional microsphere-assisted inertial microfluidics Zixiang Wang 1 & Lin Cheng 2 & Xiaoyun Wei 1 & Bo Cai 3 & Yue Sun 1 & Yuanzhen Zhang 2 & Lei Liao 1 & Xing-Zhong Zhao 1 Accepted: 13 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Being easy, safe and reliable, non-invasive prenatal diagnosis (NIPD) has been greatly pursued in recent years. Holding the complete genetic information of the fetus, fetal nucleated red blood cells (fNRBCs) are viewed as a suitable target for NIPD application. However, effective separating fNRBCs from maternal peripheral blood for clinic use still faces great challenges, given that fNRBCs are extremely rare in maternal blood circulation. Here, by combining the high-throughput inertial microfluidic chip with multifunctional microspheres as size amplification, we develop a novel method to isolate fNRBCs with high performance. To enlarge the size difference between fNRBCs and normal blood cells, we use the gelatin coated microspheres to capture fNRBCs with anti-CD147 as specific recognizer at first. The size difference between fNRBCs captured by the microspheres and normal blood cells makes it easy to purify the captured fNRBCs through the spiral microfluidic chip. Finally, the purified fNRBCs are mildly released from the microspheres by enzymatically degrading the gelatin coating. Cell capture efficiency about 81%, high purity of 83%, as well as cell release viability over 80% were achieved using spiked samples by this approach. Additionally, fNRBCs were successfully detected from peripheral blood of pregnant women with an average of 24 fNRBCs per mL, suggesting the great potential of this method for clinical non-invasive prenatal diagnosis. Keywords Non-invasiveprenatal diagnosis . Fetal nucleated red blood cell . High throughputmicrofluidic chip . Enzymatic release
1 Introduction Statistically, 3% of global newborn may occur genetic defect, most of which are chromosome abnormalities (Hui et al. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10544-020-00531-2) contains supplementary material, which is available to authorized users. * Yuanzhen Zhang [email protected] * Lei Liao [email protected] * Xing-Zhong Zhao [email protected] 1
Key Laboratory of Artificial Micro- and Nano-Structures of Ministry of Education, School of Physics and Technology, Wuhan University, Wuhan 430072, China
2
Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, People’s Republic of China
3
Research Center for Tissue Engineering and Regenerative Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
2018). In China, because of the increasing amount of pregnant women with high age and the environmental pollution, the type and the amount of inherited diseases are increasing, which have brought great burden to the society and the family
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