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Related Experiment Videos

Rapid hepatocyte spheroid formation: optimization and long-term function in perfused microcapsules

S Surapaneni1, T Pryor, M D Klein

  • 1Department of Chemical Engineering, Wayne State University, Detroit, Michigan 48202, USA.

ASAIO Journal (American Society for Artificial Internal Organs : 1992)
|November 14, 1997
PubMed
Summary

This study demonstrates a novel method for rapidly forming controlled hepatocyte spheroids, crucial for enhancing liver support systems. These engineered spheroids maintain long-term function, offering a promising advancement in bioartificial liver development.

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Area of Science:

  • Hepatocyte biology and tissue engineering
  • Biomaterials and regenerative medicine
  • Cellular engineering for bioartificial organs

Background:

  • Optimizing cell-cell interactions is vital for the long-term efficacy of liver support systems.
  • Controlling the size of hepatocyte aggregates (spheroids) is a key factor in enhancing their function.
  • Primary rat hepatocytes were utilized to investigate spheroid formation and function.

Purpose of the Study:

  • To develop a rapid and efficient protocol for inducing hepatocyte aggregation into controlled-diameter spheroids.
  • To evaluate the long-term in vitro function of microencapsulated hepatocyte spheroids.
  • To establish a method for enhancing cell-cell interactions in liver support systems.

Main Methods:

  • Primary rat hepatocytes were cultured using an intermittent settling and agitation protocol in albumin-coated flasks.

Related Experiment Videos

  • Hepatocyte suspensions were agitated at 20-minute intervals to promote spheroid formation.
  • Spheroid diameter and shape were analyzed using light microscopy and computer imaging.
  • Long-term function was assessed by microencapsulating spheroids and culturing them under perfusion for 21 days.

    • Main Results:

    • Controlled hepatocyte aggregation into spheroids with diameters ranging from 60 to 240 microns was achieved within 8-24 hours.
    • Microencapsulated spheroids demonstrated albumin secretion rates comparable to collagen sandwich controls for at least 14 days.
    • Peak albumin secretion rates reached approximately 80 microns/day/10^6 cells, particularly after medium changes.

    Conclusions:

    • A highly efficient method for controlled hepatocyte aggregation was established in under 8 hours.
    • Microencapsulated spheroids exhibit sustained in vitro liver function, indicating potential for liver support applications.
    • This technique offers a viable strategy for improving cell-cell interactions and long-term performance in bioengineered liver systems.