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Related Experiment Video

Updated: Jul 31, 2025

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Protocol for generation of multicellular spheroids through reduced gravity.

Dylan A Zinn1, Christine Mehner2, Tushar Patel3

  • 1Department of Transplantation, Mayo Clinic, Jacksonville, FL 32224, USA.

STAR Protocols
|May 3, 2023
PubMed
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This study introduces a simple method for creating multicellular spheroids using standard culture tubes and slow rotation. This technique facilitates spheroid production for drug testing and tumor biology research.

Area of Science:

  • Cell biology
  • Biotechnology
  • Drug discovery

Background:

  • Multicellular spheroids are valuable models in drug testing and tumor biology.
  • Current spheroid production methods often require specialized equipment.
  • Standardized protocols are needed for consistent and scalable spheroid generation.

Purpose of the Study:

  • To present a novel, accessible protocol for producing viable multicellular spheroids.
  • To detail methods for spheroid culture, maintenance, and expansion.
  • To enable high-throughput spheroid production using standard laboratory equipment.

Main Methods:

  • Utilized slow rotation of standard culture tubes around a horizontal axis.
  • Described detailed steps for seed and starter spheroid cultures.
Keywords:
CancerCell BiologyCell cultureCell-based AssaysOrganoids

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  • Included procedures for spheroid maintenance, expansion, and assessment (size, count, viability, immunohistochemistry).

    • Main Results:

    • Successfully produced viable multicellular spheroids using the described rotation method.
    • The protocol effectively reduces gravitational forces that cause cell clumping.
    • Demonstrated amenability of the method for high-throughput applications.

    Conclusions:

    • The presented protocol offers a simplified and effective approach to spheroid production.
    • This method is suitable for various research applications, including drug screening and cancer biology.
    • The technique is adaptable for high-throughput screening, enhancing research efficiency.