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Enrichment of Placental Trophoblast Cells from Clinical Cervical Samples Using Differences in Surface Adhesion on an Inclined Plane

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Abstract

Placental trophoblast cells present in cervical samples have great potential towards non-invasive prenatal testing. However, cervical samples are highly heterogeneous, largely comprised of maternal cervical cells with only a small quantity of trophoblast cells. In order to use these rare cells for diagnostic applications, there is a need to enrich and isolate them from the heterogeneous maternal sample. Our goal was to investigate the use of gravitational flow on an inclined surface and optimize parameters including angle of incline, surface material, incubation time on the surface, solution volume, and device channel width in order to identify a design allowing label-free enrichment of trophoblast cells. In this work we detail the development of a new method and device for controlling cell adhesion to a surface vs. rolling into a collection area. The enrichment device design was developed for ease of use by non-specialized personal and on a slide surface for the ability to be directly integrated onto an automatic cell picker instrument, which can be used for downstream single cell isolation. JEG-3 trophoblast cells were used with clinical cervical samples to present the effect of the different optimization parameters on enrichment. We further provide an assessment of the impact shear stress and thickness of the liquid layer has on cell enrichment. We found that this method provides a maximum JEG-3 enrichment using polystyrene surfaces at a 50° incline with a 5 min incubation period prior to inclined flow. This resulted in a 396 ± 52% increase in purity of the trophoblast cells from the clinical cervical samples as confirmed using human leukocyte antigen G (HLA-G) antibody staining with fluorescence imaging to identify JEG-3 cells. Ultimately, this method is inexpensive, quick, and has the potential for direct integration into fetal cell isolation platforms.

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Acknowledgments

The material is based upon work supported by the National Science Foundation Graduate Research Fellowship awarded to C.M.B.H., under Grant No. 1644760. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation. We thank the Brown Design Workshop for the use of the laser cutter and Makerbot Replicator. We thank Perkin Elmer, Inc., including Morey Kraus and Richard Joseph, for prenatal testing discussions and cervical samples.

Conflict of interest

AT is a paid scientific advisor/consultant and lecturer for PerkinElmer.

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  1. Christina M. Bailey-Hytholt and Sumaiya Sayeed are Co-first authors.

Authors and Affiliations

  1. School of Engineering, Center for Biomedical Engineering, Brown University, Providence, RI, 02912, USA

    Christina M. Bailey-Hytholt, Sumaiya Sayeed, Anita Shukla & Anubhav Tripathi

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  1. Christina M. Bailey-Hytholt
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Correspondence to Anubhav Tripathi.

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Current address for Christina M. Bailey-Hytholt: Department of Chemical Engineering, Worcester Polytechnic Institute, Worcester, MA, 01609, USA.

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Bailey-Hytholt, C.M., Sayeed, S., Shukla, A. et al. Enrichment of Placental Trophoblast Cells from Clinical Cervical Samples Using Differences in Surface Adhesion on an Inclined Plane. Ann Biomed Eng 49, 2214–2227 (2021). https://doi.org/10.1007/s10439-021-02742-x

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