Spheroid Isolation

Spheroid isolation is the process of recovering three-dimensional aggregates of cells from a culture or biological sample while preserving their structure and viability. It typically relies on gentle mechanical dissociation, selective washing, and size-based separation or low-speed centrifugation to remove single cells, debris, and excess medium without disrupting cell-cell contacts. Isolated spheroids can then be counted, transferred, embedded, dissociated, or subjected to imaging and molecular analysis. In biology, this workflow supports studies of cell adhesion, tissue organization, stem cell behavior, tumor biology, drug response, and three-dimensional models that more closely represent features of tissues than conventional monolayer cultures.

Spheroid Isolation - Related Videos

Research

JoVE Journal - Biology

Isolation of Mammary Epithelial Cells from Three-dimensional Mixed-cell Spheroid Co-culture

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Cited by 16 •

2012

A simple method is described for analyzing effects of tissue fibroblasts on associated epithelial cells. The combination of this method and three-dimensional tissue culture can facilitate analysis of cells after isolation from 3D. The technique is applicable to cells of varying malignant potential, allowing systematic study of effects of tumor-associated stroma on tumor cells.

Splitting Spheroids for Sub-culturing and Shipping: A Procedure for Propagating and Transferring Spheroids from Small Bowel Neuroendocrine Tumor

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2023

This video demonstrates a strategy to split and propagate spheroids developed from small bowel neuroendocrine tumor cells. These spheroids have small bowel neuroendocrine tumor markers and can be propagated and further used to test anti-cancer drugs.

Immunofluorescent Staining of Spheroids: A Technique to Analyze Spheroids for Expression of Intra- and Extracellular Antigen Expression

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2023

This video describes the technique of immunofluorescent staining of spheroids to understand the differentiation of the fibroblasts or CAFs. This protocol helps detect expression of intra- and extracellular antigens, including deposition of ECM proteins within the spheroids.

Embedding 3D Spheroids in Agarose Gel Drops: A Technique to Preserve Spheroids for Downstream Analysis

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2023

This video describes the technique to embed 3D spheroids in agarose gel drops for preservation. The embedded 3D spheroids can be further used for downstream analysis.

Extracellular Flux Assay in 3D Spheroids: A Method for Measuring the Metabolic Activity of Pancreatic Tumor Spheroids

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2023

This protocol describes the technique for performing extracellular flux assay on 3D spheroids generated from the co-culture of pancreatic cancer cells (Patu8902) and activated pancreatic fibroblasts (PS1) to measure the two major energy-producing pathways, glycolysis and mitochondrial respiration.

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