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Updated: Jul 3, 2025

Author Spotlight: Advancing Hematopoietic Research Using Stromal Cell Isolation for Single Cell Sequencing

Author Spotlight: Advancing Hematopoietic Research Using Stromal Cell Isolation for Single Cell Sequencing

Published on: January 26, 2024

Stromal Cell Isolation from Hematopoietic Organs.

Trine Kristiansen¹, Christina Mayerhofer¹, Karin Gustafsson²

¹Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Stem Cell Institute; Department of Stem Cell and Regenerative Biology, Harvard University.

Journal of Visualized Experiments : Jove

|February 12, 2024

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This study details protocols for isolating high-quality stromal cells from immune organs for single-cell sequencing. These methods help analyze cellular heterogeneity in health, stress, and aging.

Area of Science:

Immunology
Cell Biology
Genomics

Background:

Single-cell sequencing reveals cellular heterogeneity in hematopoietic organ stroma.
Understanding stromal cell populations is key to studying immune system function, aging, and disease.
Previous methods lacked detailed protocols for isolating specific stromal cell types.

Purpose of the Study:

To provide robust, step-wise protocols for isolating high-quality stromal cells from murine and human thymus, and murine bone marrow.
To optimize stromal cell isolation for compatibility with single-cell multiomics.
To guide researchers in achieving reproducible results in stromal cell analysis.

Main Methods:

Development of detailed protocols for stromal cell isolation from murine and human thymus and bone marrow.

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Last Updated: Jul 3, 2025

Author Spotlight: Advancing Hematopoietic Research Using Stromal Cell Isolation for Single Cell Sequencing

Author Spotlight: Advancing Hematopoietic Research Using Stromal Cell Isolation for Single Cell Sequencing

Published on: January 26, 2024

Isolation and Transplantation of Hematopoietic Stem Cells HSCs

Isolation and Transplantation of Hematopoietic Stem Cells HSCs

Published on: February 25, 2007

In Vivo Osteo-organoid Approach for Harvesting Therapeutic Hematopoietic Stem/Progenitor Cells

In Vivo Osteo-organoid Approach for Harvesting Therapeutic Hematopoietic Stem/Progenitor Cells

Published on: February 16, 2024

Optimization of sample digestion and hematopoietic lineage depletion.

Application of Fluorescence-Activated Cell Sorting (FACS) for cell purification.

Assessment of isolated cells for suitability in single-cell multiomics.

Main Results:

Established protocols yield high-quality stromal cells suitable for single-cell sequencing.
Demonstrated the impact of digestion and lineage depletion on stromal cell yield and quality.
Provided FACS profiles illustrating successful and unsuccessful dissociation techniques.
Showcased downstream stromal cell yields in post-sequencing analysis.

Conclusions:

Optimized protocols are essential for high-quality stromal cell isolation.
Careful consideration of sample processing is crucial for reproducible single-cell multiomics data.
These protocols will advance the study of stromal cell heterogeneity in hematopoietic organs.