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Secretome-Based Prediction of Three-Dimensional Hepatic Microtissue Physiological Relevance.

Amish Asthana1, Charles McRae White1, Kenneth Ndyabawe1

  • 1Cellular Bioengineering Laboratory, College of Engineering, Driftmier Engineering Center, The University of Georgia, 597 D. W. Brooks Drive, Athens, Georgia 30602, United States.

ACS Biomaterials Science & Engineering
|January 19, 2021
PubMed
Summary

Researchers identified specific cytokines, including PDGF-AB/BB and VEGF, as early biomarkers for three-dimensional (3D) cell cultures. This discovery offers a universal method to assess the complex physiological relevance (CPR) of 3D models sooner than traditional markers.

Keywords:
3D culturecell-based assayhepaticscaffoldsecretome

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Area of Science:

  • Biomarker Discovery
  • Cell Culture Technology
  • In Vitro Toxicology

Background:

  • Assessing complex physiological relevance (CPR) in three-dimensional (3D) cell models requires early biomarkers, as current indicators are detected late and necessitate diverse analytical methods.
  • Traditional markers like albumin production, CYP3A4 expression, and bile canaliculi formation are insufficient to distinguish advanced 3D cultures from conventional two-dimensional (2D) cultures.
  • A universal biomarker is needed for a standardized detection platform to evaluate 3D in vitro models.

Purpose of the Study:

  • To identify early biomarkers for assessing the complex physiological relevance (CPR) of three-dimensional (3D) cell cultures.
  • To explore the potential of the cytokine secretion profile (secretome) as an early indicator for 3D culture development.
  • To find a universal biomarker independent of cell type for a common detection platform.

Main Methods:

  • Comparative analysis of cytokine secretion profiles between 3D and 2D cell cultures.
  • Quantitative assessment of specific cytokine levels at early time points (days 3 and 4).
  • Focus on Platelet-Derived Growth Factor (PDGF-AB/BB) and Vascular Endothelial Growth Factor (VEGF) as candidate biomarkers.

Main Results:

  • Platelet-Derived Growth Factor-AB/BB (PDGF-AB/BB) was found to be upregulated in 3D cultures compared to 2D cultures at early time points.
  • Vascular Endothelial Growth Factor (VEGF) also showed upregulation in 3D cultures relative to 2D cultures during the early culture phase.
  • These specific cytokines serve as potential early indicators of 3D model development.

Conclusions:

  • The secretome, specifically PDGF-AB/BB and VEGF, presents a promising avenue for early biomarker identification in 3D cell cultures.
  • Cytokine profiling offers a more sensitive approach to differentiate 3D from 2D cultures compared to traditional hepatic markers.
  • Further in vivo validation of these identified cytokines is crucial to establish their role in predicting the complex physiological relevance of 3D in vitro models.