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연세대학교 의과대학, Emory University School of Medicine

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Circulation, November 14, 2017, Volume 136, Issue 20 | https://doi.org/10.1161/CIRCULATIONAHA.116.026329 Enhanced Therapeutic and Long-Term Dynamic Vascularization Effects of Human Pluripotent Stem Cell–Derived Endothelial Cells Encapsulated in a Nanomatrix Gel

Shin-Jeong Lee, PhD, Young-Doug Sohn, PhD, Adinarayana Andukuri, PhD, Sangsung Kim, MS, Jaemin Byun, MS, Ji Woong Han, PhD, In-Hyun Park, PhD, Ho-Wook Jun, PhD, Young-sup Yoon, MD, PhD

Division of Cardiology, Department of Medicine, Emory University School of Medicine, Atlanta, GA (S.-J.L., Y.-D.S., A.A., S.K., J.B., J.W.H., Y.-S.Y.). Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea (S.-J.L., Y.-S.Y.). Department of Genetics, Yale University School of Medicine, New Haven, CT (I.-H.P.). Department of Biomedical Engineering, University of Alabama at Birmingham (H.-W.J.).

Correspondence to: Young-sup Yoon, MD, PhD, Department of Medicine, Division of Cardiology, Emory University School of Medicine, 101 Woodruff Circle, WMB 3309, Atlanta, GA 30322

Abstract

Background: Human pluripotent stem cell (hPSC)-derived endothelial cells (ECs) have limited clinical utility because of undefined components in the differentiation system and poor cell survival in vivo. Here, we aimed to develop a fully defined and clinically compatible system to differentiate hPSCs into ECs. Furthermore, we aimed to enhance cell survival, vessel formation, and therapeutic potential by encapsulating hPSC-ECs with a peptide amphiphile (PA) nanomatrix gel.

Methods: We induced differentiation of hPSCs into the mesodermal lineage by culturing on collagen-coated plates with a glycogen synthase kinase 3β inhibitor. Next, vascular endothelial growth factor, endothelial growth factor, and basic fibroblast growth factor were added for endothelial lineage differentiation, followed by sorting for CDH5 (VE-cadherin). We constructed an extracellular matrix-mimicking PA nanomatrix gel (PA-RGDS) by incorporating the cell adhesive ligand Arg-Gly-Asp-Ser (RGDS) and a matrix metalloproteinase-2-degradable sequence. We then evaluated whether the encapsulation of hPSC-CDH5⁺ cells in PA-RGDS could enhance long-term cell survival and vascular regenerative effects in a hind-limb ischemia model with laser Doppler perfusion imaging, bioluminescence imaging, real-time reverse transcription-polymerase chain reaction, and histological analysis.

Results: The resultant hPSC-derived CDH5⁺ cells (hPSC-ECs) showed highly enriched and genuine EC characteristics and proangiogenic activities. When injected into ischemic hind limbs, hPSC-ECs showed better perfusion recovery and higher vessel-forming capacity compared with media-, PA-RGDS-, or human umbilical vein EC-injected groups. However, the group receiving the PA-RGDS-encapsulated hPSC-ECs showed better perfusion recovery, more robust and longer cell survival (> 10 months), and higher and prolonged angiogenic and vascular incorporation capabilities than the bare hPSC-EC-injected group. Surprisingly, the engrafted hPSC-ECs demonstrated previously unknown sustained and dynamic vessel-forming behavior: initial perivascular concentration, a guiding role for new vessel formation, and progressive incorporation into the vessels over 10 months.

Conclusions: We generated highly enriched hPSC-ECs via a clinically compatible system. Furthermore, this study demonstrated that a biocompatible PA-RGDS nanomatrix gel substantially improved long-term survival of hPSC-ECs in an ischemic environment and improved neovascularization effects of hPSC-ECs via prolonged and unique angiogenic and vessel-forming properties. This PA-RGDS-mediated transplantation of hPSC-ECs can serve as a novel platform for cell-based therapy and investigation of long-term behavior of hPSC-ECs.

endothelial cells, pluripotent stem cells, regeneration, stem cell, vascular diseases

논문정보

형식Research article
게재일2017년 11월 (BRIC 등록일 2017-11-22)
연구진국내(교신)+국외 연구진
분야 의약학 > 재생의학

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