Emulsion-based encapsulation of pluripotent stem cells in hydrogel microspheres for cardiac differentiation.
Samuel ChangFerdous FinkleaBianca WilliamsHanna HammonsAlexander HodgeSamantha ScottElizabeth A LipkePublished in: Biotechnology progress (2020)
Cardiovascular disease is the leading cause of death worldwide, and current treatments are ineffective or unavailable to majority of patients. Engineered cardiac tissue (ECT) is a promising treatment to restore function to the damaged myocardium; however, for these treatments to become a reality, tissue fabrication must be amenable to scalable production and be used in suspension culture. Here, we have developed a low-cost and scalable emulsion-based method for producing ECT microspheres from poly(ethylene glycol) (PEG)-fibrinogen encapsulated mouse embryonic stem cells (mESCs). Cell-laden microspheres were formed via water-in-oil emulsification; encapsulation occurred by suspending the cells in hydrogel precursor solution at cell densities from 5 to 60 million cells/ml, adding to mineral oil and vortexing. Microsphere diameters ranged from 30 to 570 μm; size variability was decreased by the addition of 2% poly(ethylene glycol) diacrylate. Initial cell encapsulation density impacted the ability for mESCs to grow and differentiate, with the greatest success occurring at higher cell densities. Microspheres differentiated into dense spheroidal ECTs with spontaneous contractions occurring as early as Day 10 of cardiac differentiation; furthermore, these ECT microspheres exhibited appropriate temporal changes in gene expression and response to pharmacological stimuli. These results demonstrate the ability to use an emulsion approach to encapsulate pluripotent stem cells for use in microsphere-based cardiac differentiation.
Keyphrases
- gene expression
- single cell
- cardiovascular disease
- cell therapy
- low cost
- left ventricular
- drug delivery
- end stage renal disease
- chronic kidney disease
- type diabetes
- pluripotent stem cells
- molecularly imprinted
- stem cells
- cell cycle arrest
- fatty acid
- embryonic stem cells
- coronary artery disease
- ejection fraction
- metabolic syndrome
- mesenchymal stem cells
- high resolution
- cell death
- mass spectrometry
- bone marrow
- oxidative stress
- tissue engineering
- atrial fibrillation