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Unlocking Bio-Instructive Polymers: A Novel Multi-Well Screening Platform Based on Secretome Sampling.

Shirin Fateh1, Reem A Alromaihi2, Amir M Ghaemmaghami2

  • 1School of Pharmacy, University of Nottingham, Nottingham, UK.

Bio-Protocol
|February 26, 2024
PubMed
Summary

This study introduces a novel multi-well platform for high-throughput screening of biomaterials, enabling rapid secretome profiling of both adherent and non-adherent cells for improved biocompatibility assessment.

Keywords:
2D printingBio-instructiveBiomaterialsHTSHigh-throughput screeningImmune-instructivePolymer microarraySecretome profiling

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • High-Throughput Screening

Background:

  • Traditional biomaterial development faces challenges due to limited understanding of cell-material interactions and adverse biological responses.
  • Existing high-throughput screening (HTS) platforms often lack the capability to analyze non-adherent cells or their secreted factors.
  • Secreted proteins (secretome) play a crucial role in regulating cell behavior and determining the success of implanted biomaterials.

Purpose of the Study:

  • To develop and validate a versatile multi-well platform for high-throughput screening of biomaterials.
  • To enable secretome profiling of both adherent and non-adherent cells cultured on printed polymer surfaces.
  • To facilitate the discovery of novel biomaterials with enhanced biocompatibility and predictable cellular responses.

Main Methods:

  • Development of a multi-well platform with individual cell culture chambers compatible with robotic polymer printing.
  • Preparation of polymer substrates including acrylate-based polymers via O2 plasma etching, functionalization, and Poly(2-hydroxyethyl methacrylate) (pHEMA) dip coating.
  • Validation of the platform using adherent (macrophages) and non-adherent (dendritic cells) primary human cells, followed by secretome analysis via ELISA for cytokines.

Main Results:

  • Successful fabrication of a multi-well platform enabling robotic printing of 7 mm x 7 mm polymer surfaces within confined areas.
  • Demonstrated compatibility of the platform with both adherent and non-adherent primary human cells.
  • Showcased the platform's utility for secretome analysis, detecting multiple cytokines secreted by cultured cells.

Conclusions:

  • The developed multi-well platform offers a robust solution for high-throughput biomaterial screening, overcoming limitations of existing methods.
  • This platform facilitates comprehensive analysis of cell-material interactions by incorporating secretome profiling for diverse cell types.
  • It is expected to accelerate the discovery of advanced biomaterials and broaden their applications by integrating cell-bound and secreted factor assessments.