Engineering surfaces for site-specific vascular differentiation of mouse embryonic stem cells |
| |
| Authors: | C. Katherine Chiang Mohammad Fahad Chowdhury Rohin K. Iyer William L. Stanford Milica Radisic |
| |
| Affiliation: | 1. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Ont., Canada;2. Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ont., Canada;3. Ontario Human iPS Cell Facility, University of Toronto, Ont., Canada;1. Key Laboratory of Green Processing and Functional Textiles of New Textile Materials, Ministry of Education, Wuhan Textile University, Wuhan 430073, People’s Republic of China;2. Research School of Chemistry, Australian National University, Canberra, ACT 2601, Australia;1. Department of Orthopedic Surgery, Stanford University, Stanford, CA, USA;2. Extremity Trauma & Regenerative Medicine Task Area, US Army Institute of Surgical Research, San Antonio, TX, USA;3. Department of Craniomaxillofacial Regenerative Medicine, Dental and Trauma Research Detachment, United States Army Institute of Surgical Research, Fort Sam Houston, TX, USA;4. Department of Prosthodontics, Dental Science Research Institute and BK21 Project, School of Dentistry, Chonnam National University, Gwangju, Republic of Korea;5. Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA;1. Division of Plastic Surgery, Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan;2. Institute of Clinical Medicine, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan;3. Department of Cell Biology and Anatomy, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan;4. Institute of Basic Medical Sciences, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan;5. Department of Occupational Therapy, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan;6. Department of Occupational Therapy, I-Shou University, No.1, Sec.1, Syuecheng Rd., Dashu District, Kaouhsiung City 84001, Taiwan;7. Department of Biomedical Engineering, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan;8. Medical Device Innovation Center, National Cheng Kung University, No. 1, University Rd., Tainan 701, Taiwan |
| |
| Abstract: | Differentiation of stem and progenitor cells routinely relies on the application of soluble growth factors, an approach that enables temporal control of cell fate but enables no spatial control of the differentiation process. Angiogenic progenitor cells derived from mouse embryonic stem cells (ESCs) were differentiated here according to the pattern of immobilized vascular endothelial growth factor-A (VEGF). Mouse ESCs engineered to express green fluorescent protein (eGFP) under control of promoter for the receptor tyrosine kinase Flk1 were used. The Flk1+ angiogenic progenitors were selected from day 3 differentiating embryoid bodies based on their expression of eGFP using fluorescence activated cell sorting. Mouse VEGF165 was covalently immobilized onto collagen IV (ColIV) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) chemistry. A non-cell adhesive layer of photocrosslinkable chitosan was first created, after which VEGF–ColIV was stamped as 100 μm wide lanes on top of the chitosan layer and the Flk1+ angiogenic progenitors were seeded for site-specific differentiation. Lanes stamped with only ColIV served as controls. The results presented here demonstrate that the cultivation of Flk1+ progenitors on surfaces with immobilized VEGF yielded primarily endothelial cells (53 ± 13% CD31 positive and 17 ± 2% smooth muscle actin positive), whereas surfaces without VEGF favored vascular smooth muscle-like cell differentiation (26 ± 17% CD31 positive and 38 ± 9% smooth muscle actin positive). |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
