Utilizing An Induced Pluripotent Stem Cells to Model Varicose Vein
Chikage Noishiki, MD, PhD, Shaunak S. Adkar, MD, PhD, David Wu, BS, Lu Liu, PhD, Naima C. Turbes, BS, Dipti Tripathi, PhD, Amit Manhas, PhD, Dilip Thomas, PhD, Karim Sallam, MD, Jason T. Lee, MD, Nicholas J. Leeper, MD, Derek M. Klarin, MD, Eri Fukaya MD, PhD, Nazish Sayed, MD, PhD
Introduction: Varicose veins (VV) are a multifactorial vascular disorder influenced by genetic and epigenetic factors. The disease is characterized by venous hypertension, inflammation, and vein wall dilation, affecting endothelial cells (ECs) and smooth muscle cells (SMCs). However, due to limited access to patient-derived ECs and SMCs, the molecular mechanisms underlying VV remain poorly understood. Induced pluripotent stem cells (iPSCs) provide a promising approach to model disease phenotypes in vitro, enabling the generation of patient-specific vascular cells while preserving the genetic background of the donor.
Methods: We recruited 50 VV patients from our Stanford clinic and isolated peripheral blood mononuclear cells (PBMCs) for reprogramming into iPSCs. These iPSCs were then differentiated into ECs using a chemically defined monolayer differentiation protocol. The differentiation efficiency was assessed by flow cytometry for CD31 expression, and RT-qPCR was performed to evaluate the expression of endothelial markers PECAM1, NOS3, CDH5, and VWF.
Results: Our differentiation protocol consistently yielded 80–95% CD31+ iPSC-ECs. RT-qPCR confirmed the expression of key endothelial markers in both control and VV-derived ECs, demonstrating successful differentiation and providing a platform to study VV-specific cellular characteristics.
Conclusion: This patient-specific iPSC model allows for the investigation of individual responses to treatments and identification of genetic variants contributing to VV. However, a clear distinction between venous and arterial phenotypes was not assessed. Future studies will focus on characterizing the functional and developmental differences between venous and arterial iPSC-ECs to further elucidate VV pathogenesis.