Electric-Pulse Current Stimulation Increases If Current in mShox2 Genetically Modified Canine Mesenchymal Stem Cells

Area of Science:

• Cardiovascular Research
• Stem Cell Biology
• Molecular Cardiology

Background:

• The If pacemaker current is crucial for cardiac automaticity.
• Understanding the genetic and electrical factors influencing pacemaker generation is vital for regenerative medicine.

Purpose of the Study:

• To investigate the role of mShox2 in generating the If pacemaker current in canine mesenchymal stem cells (cMSCs).
• To evaluate the effect of electric-pulse current stimulation (EPCS) on mShox2-modified cMSCs differentiation into pacemaker-like cells.

Main Methods:

• Genetic modification of cMSCs with mShox2 and red fluorescent protein.
• Patch clamp analysis to characterize the generated inward current.
• qRT-PCR and Western blotting to assess gene and protein expression of cardiac markers.
• Morphological assessment and detection of cardiomyogenic differentiation marker cTnT.

Main Results:

• mShox2 modification and EPCS induced a time- and voltage-dependent inward current, identified as If current.
• EPCS significantly increased the detection rate, amplitude, and density of If current.
• Upregulation of pacemaker markers (Tbx3, HCN4, Cx45) and downregulation of working myocardium markers (Nkx2.5, Cx43) were observed.
• Cells exhibited morphological changes consistent with cardiomyogenic differentiation, including cTnT expression.

Conclusions:

• EPCS promotes the differentiation of mShox2-modified cMSCs into pacemaker-like cells.
• This differentiation leads to enhanced generation of the If pacemaker current.