Section of Plastic Surgery, Department of Surgery, Yale School of Medicine, Yale University, New Haven, Connecticut, USA.
Department of Biomedical Engineering, Yale University, New Haven, Connecticut, USA.
J Biomed Mater Res A. 2022 Nov;110(11):1813-1823. doi: 10.1002/jbm.a.37423. Epub 2022 Jul 11.
Human-induced pluripotent stem cells (hiPSC) and their differentiated vascular cells have been revolutionizing the field of regenerative wound healing. These cells are shown to be rejuvenated with immense potentials in secreting paracrine factors. Recently, hiPSC-derived vascular smooth muscle cells (hiPSC-VSMC) have shown regenerative wound healing ability via their paracrine secretion. The quest to modulate the secretory function of these hiPSC-VSMC is an ongoing effort and involves the use of both biochemical and biophysical stimuli. This study explores the development and optimization of a reproducible, inexpensive protocol to form hiPSC-VSMC derived spheroids to investigate the implications of spheroid size on viability and paracrine secretion. Our data show the successful formation of different sizes of spheroids using various amount of hiPSC-VSMC. The hiPSC-VSMC spheroids formed with 10,000 cells strike an ideal balance between overall cell health and maximal paracrine secretion. The conditioned medium from these spheroids was found to be bioactive in enhancing human dermal fibroblast cell proliferation and migration. This research will inform future studies on the optimal spheroid size for regenerative wound healing applications.
人诱导多能干细胞（hiPSC）及其分化的血管细胞正在彻底改变再生性伤口愈合领域。这些细胞显示出通过旁分泌因子分泌具有巨大潜力的年轻化。最近，hiPSC 衍生的血管平滑肌细胞（hiPSC-VSMC）通过旁分泌分泌显示出再生性伤口愈合能力。调节这些 hiPSC-VSMC 的分泌功能是一项正在进行的努力，涉及使用生化和生物物理刺激。本研究探讨了开发和优化一种可重复、低成本的方案，以形成 hiPSC-VSMC 衍生的球体，以研究球体大小对活力和旁分泌分泌的影响。我们的数据显示，使用不同数量的 hiPSC-VSMC 成功形成了不同大小的球体。用 10,000 个细胞形成的 hiPSC-VSMC 球体在整体细胞健康和最大旁分泌分泌之间达到了理想的平衡。这些球体的条件培养基被发现具有生物活性，可增强人真皮成纤维细胞的增殖和迁移。这项研究将为再生性伤口愈合应用的最佳球体大小提供信息。
