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High-throughput isolation of fetal nucleated red blood cells by multifunctional microsphere-assisted inertial microfluidics

Biomedical Microdevices volume 22, Article number: 75 (2020) Cite this article

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.

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Acknowledgements

Authors Z. X. Wang and L. Cheng contributed equally to this work. This work is supported by the National R&D Program for Major Research Instruments (No. 81527801), the National Natural Science Foundation of China (No. 51272184, 81572860, and 61474084) and the National Science Fund for Talent Training in Basic Science (No. J1210061). National Key R&D Program (Grant No. 2016YFC1000701).

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Affiliations

  1. Key Laboratory of Artificial Micro- and Nano-Structures of Ministry of Education, School of Physics and Technology, Wuhan University, Wuhan, 430072, China

    Zixiang Wang, Xiaoyun Wei, Yue Sun, Lei Liao & Xing-Zhong Zhao

  2. Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, 430071, People’s Republic of China

    Lin Cheng & Yuanzhen Zhang

  3. Research Center for Tissue Engineering and Regenerative Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Bo Cai

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  1. Zixiang Wang
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  2. Lin Cheng
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  3. Xiaoyun Wei
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  5. Yue Sun
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  6. Yuanzhen Zhang
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  7. Lei Liao
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  8. Xing-Zhong Zhao
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Correspondence to Yuanzhen Zhang, Lei Liao or Xing-Zhong Zhao.

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Wang, Z., Cheng, L., Wei, X. et al. High-throughput isolation of fetal nucleated red blood cells by multifunctional microsphere-assisted inertial microfluidics. Biomed Microdevices 22, 75 (2020). https://doi.org/10.1007/s10544-020-00531-2

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Keywords

  • Non-invasive prenatal diagnosis
  • Fetal nucleated red blood cell
  • High throughput microfluidic chip
  • Enzymatic release