Spontaneous cardiomyocyte differentiation of mouse embryoid bodies regulated by hydrogel crosslink density
Open accessAbstract
Cellular therapies have great potential to provide alternative treatment options for those suffering from heart disease. In order to optimize cell delivery for therapeutic efficacy, a greater understanding of parameters that impact stem cell differentiation, survival, growth, and development are needed. In this study, we examine the role of hydrogel crosslink density on spontaneous cardiomyocyte (CM) differentiation of murine embryoid bodies (EBs). CM differentiation was accelerated in hydrogels of low crosslink density, where 100% of the hydrogels were positive for CM differentiation compared to only 53% in the high crosslink density group after 8 days of culture. DNA microarray data suggests that enhanced CM differentiation in the low crosslink density hydrogels was not tissue specific but rather a result of favoured EB development and cell proliferation. Additionally, enhanced EB growth and differentiation in low crosslink density hydrogels was independent of RGD ligand density and not a consequence of enhanced diffusion. We also demonstrate that matrix metalloproteinase activity is required for spontaneous CM differentiation in 3D hydrogels. Low hydrogel crosslink density regulates spontaneous EB differentiation by promoting EB growth and development. Elucidating the effects of microenvironmental cues on cell differentiation can aid in the optimization of stem cell-based therapies for tissue regeneration.
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Among the elastic moduli studied (700, 3000, and 4000 Pa), the softest material favored the proliferation of embryoid bodies that contain mesodermal progenitor cells and promoted rapid cardiomyocyte differentiation.
“Among the elastic moduli studied (700, 3000, and 4000 Pa), the softest material favored the proliferation of embryoid bodies that contain mesodermal progenitor cells and promoted rapid cardiomyocyte differentiation (Fig. 5).”
From Towards organoid culture without Matrigel (2021) · cites this paper as [190]
Embryoid bodies cultured in the 700 Pa matrix displayed the highest level of MMP secretion.
“Embryoid bodies cultured in the 700 Pa matrix displayed the highest level of MMP secretion.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [190]
Murine cardiomyocyte differentiation (measured by α-myosin expression, cell contractility, and metabolic activity) was found to be dependent on the stoichiometric ratio of THPC to protein, which tuned the stiffness of the hydrogel.
“In a follow-up study, murine cardiomyocyte differentiation (measured by α-myosin expression, cell contractility, and metabolic activity) was found to be dependent on the stoichiometric ratio of THPC to protein, which tuned the stiffness of the hydrogel.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [190]
Inhibition of MMP secretion was deleterious to proliferation and differentiation, suggesting that remodeling of the matrix is essential in cardiomyocyte differentiation.
“Inhibition of MMP secretion was deleterious to proliferation and differentiation, suggesting that remodeling of the matrix is essential in cardiomyocyte differentiation190.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [190]
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- Authors
- Chung, C., Pruitt, B. L., & Heilshorn, S. C.
- Affiliations
- Stanford University (US)
- Corresponding author
- Heilshorn, Sarah C.
- Venue
- Biomaterials Science
- Publisher
- Royal Society of Chemistry (RSC)
- Published
- 2013
- Volume
- 1
- Issue
- 10
- Pages
- 1082
- ISSN
- 2047-4830 (ISSN-L)
- 2047-4849
- PMC
- PMC3987919
- OpenAlex
- W2042068815
- Cited by
- 34
- References
- 44
- Citations by year
- 2026: 1 · 2024: 1 · 2023: 3 · 2022: 5 · 2021: 2 · 2020: 6 · 2019: 2 · 2018: 3 · 2017: 1 · 2016: 4 · 2015: 2 · 2014: 3 · 2013: 1
- FWCI
- 1.53
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Biomaterials Science (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- National Institutes of Health (5r21ar062359-02, 1dp2od006477-01)
- Directorate for Biological Sciences
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK085720)
- National Institute of Arthritis and Musculoskeletal and Skin Diseases (R21-AR062359)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Keywords
- cardiomyocyte differentiation, embryoid bodies, 3D hydrogels, matrix metalloproteinase activity, stem cell therapy, heart disease, cell proliferation, microenvironmental cues, tissue regeneration, cell delivery
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