Engineered Human Adipose-Derived Stem Cells Inducing Endothelial Lineage and Angiogenic Response

• Published in: Tissue engineering. Part C, Methods Methods, 2019-03, Vol.25 (3), p.148-159
• Main Authors: Ajit, Amita, Santhosh Kumar, T.R., Krishnan, Lissy K.
• Format: Article
• Language: English
• Subjects: Angiogenesis; Bioavailability; Bioengineering; Cell culture; Cell growth; Cell Lineage; Cell migration; Cell Proliferation; Cell viability; Cells, Cultured; Cytomegalovirus; Endothelial cells; Endothelium, Vascular - cytology; Endothelium, Vascular - physiology; Experiments; Gene expression; Human Umbilical Vein Endothelial Cells; Humans; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Hypoxia-inducible factors; Medical research; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - metabolism; Mesenchymal Stem Cells - physiology; Mesenchyme; Methods Articles; Neovascularization, Physiologic; Paracrine signalling; Phenotypes; Regenerative medicine; Safety; Secretome; Stem cells; Stromal cells; Thrombosis; Tissue engineering; Tissue Engineering - methods; Transcription; Transfection; Translation; Umbilical vein; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism; Vascular endothelial growth factor receptor 2; Vascularization; Vehicles; Wound healing
• ISSN: 1937-3384; 1937-3392
• DOI: 10.1089/ten.tec.2018.0333

Biotechnological interventions to engineer favorable biochemical environments that facilitate wound repair through quick attainment of vasculature are of immense demand. Potential of angiogenic factors (AF), notably vascular endothelial growth factor A (VEGF-A) and hypoxia-inducible factor-1α (HIF-1α) and the paracrine role of human adipose-derived mesenchymal stromal cells (hADMSCs) in establishing vasculature has been reported. Independent application of AF or hADMSCs has led to clinical failure, pertaining to poor means of delivery, deprived bioavailability, and insufficiency to drive adequate angiogenesis, respectively. Although augmenting well-timed release of AF through bioengineered hADMSCs seems appealing, effective nonviral transfection in primary hADMSCs continue to be underexplored in the context of quick reproducibility and translational safety. Based on high safety and clinical value, Neon ® Transfection System in primary hADMSCs transfection was explored in this study. Multiple donor cell experiments established ∼50% transfection efficiency, acceptable cell viability, and posttransfection phenotype maintenance with safe and transient AF delivery. The delivered AF induced endothelial lineage commitment of engineered/nonengineered heterogeneous hADMSCs in culture. Delivered AF was established to function in a dose-dependent manner in terms of effect on human umbilical vein endothelial cell proliferation, migration, tube elongation, and elevated Flk-1 expression, emphasizing combinatorial AF therapy to be superior. This in vitro study is the first to report Neon Transfection System for reproducible bioengineering of primary hADMSCs for functional AF (VEGF-A & HIF-1α) delivery, demonstrating endothelial lineage commitment, predicting combinatorial effectiveness and translational safety for subsequent regenerative medicine application.