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Production of Elastin-like Protein Hydrogels for Encapsulation and Immunostaining of Cells in 3D
Published on: May 19, 2018
Characterizing ECM production by cells encapsulated in hydrogels.
Iossif A Strehin1, Jennifer H Elisseeff
1Department of Biomedical Engineering, John Hopkins University, Baltimore, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 28, 2009
Summary
This study details methods for assessing cell-produced extracellular matrix (ECM) within hydrogel scaffolds. It covers quantifying collagen, glycosaminoglycan (GAG), and DNA, alongside gene expression and histological analyses for tissue engineering.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Hydrogels, derived from polymers like polyethylene glycol and alginate, are utilized as scaffolds in tissue engineering.
- Characterizing the extracellular matrix (ECM) produced by encapsulated cells is crucial for evaluating scaffold performance.
Purpose of the Study:
- To describe detailed procedures for encapsulating cells within hydrogel scaffolds.
- To outline methods for characterizing the resultant extracellular matrix (ECM) production.
- To provide protocols for biochemical, gene expression, and histological analyses of ECM.
Main Methods:
- Cell encapsulation within polyethylene glycol and alginate hydrogels.
- Biochemical assays to quantify collagen, glycosaminoglycan (GAG), and DNA content.
- Gene expression analysis for ECM molecules (collagen I, collagen II, aggrecan).
- Histological staining (Hematoxylin and Eosin, Safranin-O) for matrix production and GAG synthesis.
Main Results:
- Quantification of key ECM components (collagen, GAG, DNA) within hydrogel scaffolds.
- Assessment of specific ECM gene expression levels.
- Histological evaluation of overall matrix deposition and GAG distribution.
Conclusions:
- Established protocols enable comprehensive characterization of cell-mediated ECM production in hydrogels.
- These methods are valuable for assessing the efficacy of hydrogel scaffolds in tissue engineering.
- The described techniques can be adapted for evaluating other scaffold materials beyond hydrogels.
