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Enhanced Proliferation of Endothelial Progenitor Cells Post-Ultrasonic Microbubble Transfection Is Plasmid DNA Size
Yi-Nan Lee1, Chao-Feng Lin2, Chiung-Yin Chang1
1Cardiovascular Center, Departments of Internal Medicine and Medical Research, Mackay Memorial Hospital, Taipei City, Taiwan.
Ultrasound in Medicine & Biology
|June 29, 2019
Summary
Ultrasonic microbubble transfection (UMT) effectively delivers large DNA plasmids into endothelial progenitor cells (EPCs). This method enhances cell proliferation by increasing interleukin-6 (IL-6) secretion, particularly with 5.6 and 9.2 kb plasmids.
Area of Science:
- Gene delivery and cell engineering
- Biotechnology and biomedical engineering
Background:
- Efficient gene delivery into endothelial progenitor cells (EPCs) is crucial for regenerative medicine.
- Plasmid size can influence transfection efficiency and subsequent biological effects.
Purpose of the Study:
- To evaluate the efficacy of ultrasonic microbubble transfection (UMT) for delivering large luciferase plasmids (5.6, 9.2, and 33 kb) into porcine EPCs.
- To assess the impact of UMT-mediated gene delivery on EPC proliferation and the secretion of key signaling molecules.
Main Methods:
- Porcine venous blood EPCs were cultured with plasmid DNA (pDNA) of varying sizes.
- Ultrasonic microbubble transfection (UMT) was applied to facilitate pDNA delivery.
- Real-time PCR and functional assays were performed to analyze gene expression and cellular responses, including proliferation and cytokine secretion.
Main Results:
- UMT enhanced luciferase expression, though efficiency decreased with increasing plasmid size (5.6 and 9.2 kb showed better results than 33 kb).
- Transfection of 5.6 and 9.2 kb pDNAs significantly enhanced EPC proliferation.
- Interleukin-6 (IL-6) secretion increased in EPCs transfected with 5.6 and 9.2 kb pDNAs; blocking IL-6 neutralized the enhanced proliferation.
Conclusions:
- UMT is an effective method for transfecting EPCs with large pDNAs, with optimal results for 5.6 and 9.2 kb plasmids.
- UMT-mediated delivery of 5.6 and 9.2 kb pDNAs stimulates EPC proliferation via increased IL-6 secretion.
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