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Chondrogenesis of Adipose-Derived Stem Cells Using an Arrayed Spheroid Format.
Robert A Gutierrez1, Vera C Fonseca2, Eric M Darling1,2,3,4
1Center for Biomedical Engineering, Brown University, Box G-B397, Providence, RI 02912 USA.
Cellular and Molecular Bioengineering
|December 19, 2022
Summary
Microscale spheroid cultures enable high-throughput assessment of adipose-derived stem cells (ASCs) chondrogenesis. This method uses fewer cells and reagents than traditional micromass cultures, proving effective for drug screening and tissue repair research.
Area of Science:
- Stem cell biology
- Tissue engineering
- Biomaterials science
Background:
- Adipose-derived stem cells (ASCs) are crucial for regenerative medicine.
- Assessing ASC chondrogenesis traditionally requires large cell numbers and volumes, hindering high-throughput applications.
- Microscale spheroid cultures offer a promising alternative for efficient cell-based assays.
Purpose of the Study:
- To evaluate the efficacy of microscale spheroid cultures for assessing ASC chondrogenesis.
- To compare chondrogenic responses in spheroid cultures versus traditional micromass cultures.
- To establish a scalable platform for high-throughput screening of chondrogenic differentiation.
Main Methods:
- Fabrication of agarose microwells for spheroid formation.
- Seeding ASCs at various densities (50 to 250,000 cells) in microwells, well plates, and centrifuge tubes.
- Culture of cells in control or chondrogenic media for 21 days with regular media changes.
Main Results:
- ASCs successfully formed viable spheroids/micromasses in all tested conditions.
- Chondrogenic conditions promoted more robust cell growth and differentiation.
- Key chondrogenic markers (sulfated glycosaminoglycans, collagen II) were detected in 5000-cell spheroids and 250,000-cell micromasses.
- Collagen I deposition, indicative of fibrocartilage, was higher in large micromasses than in small spheroids.
Conclusions:
- High-throughput spheroid formats consistently induce chondrogenic differentiation in ASCs.
- Seeding 5000 cells per spheroid is an effective density for chondrogenesis studies.
- This microscale approach facilitates high-throughput screening for tissue repair and drug-induced remodeling studies.
