简介：
Overview
This protocol describes the generation of highly pure sinus nodal cells from murine pluripotent stem cells. The process involves inducing differentiation, forming embryonic bodies, and selecting cardiomyocytes through antibiotic treatment.
Key Study Components
Area of Science
- Cardiac biology
- Stem cell research
- Cell differentiation
Background
- Sinus nodal tissue is crucial for heart rhythm regulation.
- Pluripotent stem cells can differentiate into various cell types.
- Antibiotic selection can enhance the purity of desired cell types.
- Live cell imaging is essential for assessing cell functionality.
Purpose of Study
- To produce functional sinus nodal tissue from PSC.
- To achieve a high purity of pacemaker cells.
- To demonstrate the ability of these cells to pace myocardium ex vivo.
Methods Used
- Induction of cell differentiation through medium modification.
- Formation of embryonic bodies using hanging drops.
- Seating embryonic bodies on gelatin-coated dishes.
- Antibiotic selection to isolate cardiomyocytes.
Main Results
- Generation of “Induced-sinoatrial-bodies” with over 80% pacemaker cells.
- Increased beating rates observed in the generated cells.
- Ability of iSABs to pace myocardium ex vivo confirmed.
Conclusions
- The protocol successfully generates pure sinus nodal cells.
- High purity and functionality of cells are critical for cardiac research.
- This method can advance studies in cardiac tissue engineering.
What are induced-sinoatrial-bodies?
Induced-sinoatrial-bodies are aggregates of pacemaker cells derived from pluripotent stem cells.
How is cell differentiation induced?
Cell differentiation is induced by modifying the culture medium and using hanging drops to form embryonic bodies.
What role does antibiotic selection play?
Antibiotic selection is used to ensure that only cardiomyocytes survive, enhancing the purity of the cell population.
How are the beating rates of cells assessed?
Live cell imaging is utilized to monitor and confirm the high beating frequencies of the sinus nodal cells.
What is the significance of generating pure sinus nodal cells?
Pure sinus nodal cells are essential for understanding heart rhythm and developing cardiac therapies.
Can these cells pace myocardium?
Yes, the generated cells have been shown to effectively pace myocardium in ex vivo experiments.
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