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Engineering and Validation of Open Microfluidic Platform for Organ on a Chip

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dc.contributor.advisor전누리-
dc.contributor.author탁동하-
dc.date.accessioned2018-05-28T16:07:01Z-
dc.date.available2018-05-28T16:07:01Z-
dc.date.issued2018-02-
dc.identifier.other000000151442-
dc.identifier.urihttps://hdl.handle.net/10371/140554-
dc.description학위논문 (박사)-- 서울대학교 대학원 : 공과대학 기계항공공학부, 2018. 2. 전누리.-
dc.description.abstractOpen microfluidic device was developed using the open-access microfluidic device with through-hole membrane for the regenerative medicine of Peripheral arterial disease (PAD). PAD is commonly defined as narrowing of blood vessels in the lower part of arteries or arterioles. Major risk factors of occurrence of PAD include smoking, hypertension, hypercholesterolemia, atherosclerosis, and complications of diabetes. To develop safe and effective procedures for vascularized tissue injection, we focused on open microfluidic device using through-hole membrane and open-access microfluidic device system. For the transplantable tissue culture, simple fabrication method of through-hole membrane was developed for the media supply to the tissue. Due to the small scale of the fabricated pores, the construction of through-hole membranes on a large scale and with relatively large areas faces many difficulties. Novel fabrication methods for a large-area, freestanding micro/nano through-hole membrane constructed from versatile membrane materials using through-hole membranes on a microfluidic chip (THMMC) for the reconstitution of 3D tissue. The through-hole site was easily customizable from the micro to the nanoscale, with a low or high aspect ratio giving rise to reliable membranes. Also, the rigidity and biocompatibility of the through-hole membrane are easily tunable by simple injection of versatile membrane materials to obtain a large area (up to 3600 mm2). And we describe a simple, versatile method of generating open-access microfluidic device (OAMD) with possible non-destructive tissue sampling for TEM imaging. Generally, the analysis of organ-on-a-chip usually applied by optical microscope, fluorescence microscope and confocal microscopy. Although optical imaging technologies are widespread and effective observational tools, they possess functional and resolution limitations. The myelination by Schwann cells is critically important in restoring neuromuscular motor function after injury or peripheral neuropathy, and in the case of quantifying myelination, transmission electron microscope (TEM) analysis is a requisite. The proposed OAMD platform incorporated a novel biocompatible self-detachable photopolymer (BSP) substrate to provide a viable closed microphysiological system culture environment while also allowing for controllable and nondestructive tissue sampling for TEM analysis. Furthermore, We herein thesis a novel transplantable tissue engineering technique that yields functional and vascularized tissue that can be successfully transplanted into the Balb C Nu nude mouse using THMMC and OAMD technology.-
dc.description.tableofcontentsChapter 1. Introduction 1
1-1. Open Microfluidic Platform 1
1-2. Novel Fabrication Process of Through-hole Membrane 3
1-3. Open-Access Microfluidic Device for TEM Analysis 6
1-4. Transplantation of 3D Tissue as Regenerative Medicine 8
Chapter 2. Rapid large area fabrication of multiscale through-hole membranes 10
2-1. Introduction 10
2-2. Materials and Method 11
2-2-1. Through-Hole Membrane by Microfluidic Chip (THMMC) 11
2-2-2. Distribute Microfluidic Channel 13
2-2-3. Micro / Nano Membrane 13
2-3. Result and Discussion 15
2-3-1. Microscale Through-hole Membrane 15
2-3-2. Nanoscale Through-hole Membrane 24
2-4. Conclusion 27
Chapter 3. Open-access microfluidic device (OAMD) for TEM analysis of 3D Reconstituted myelin sheaths 28
3-1. Introduction 28
3-2. Materials and Method 34
3-2-1. Biocompatible and Self-detachable Photopolymer 34
3-2-2. Open-Access Microfluidic Device 37
3-2-3. Contact Angle Measurements 37
3-2-4. Scanning Electron Microscope 38
3-2-5. Swelling Ratio 38
3-2-6. Nano Indentation 38
3-2-7. Live Dead Assay 39
3-2-8 Energy-dispersive X-ray spectroscopy (EDXS) 39
3-2-9 Universal Testing Machine (UTM) 39
3-2-10.MN-SC Coculture on a Microfluidic Chip 40
3-2-11. ICC: Immunocytochemistry 43
3-2-12. Western Blotting 43
3-2-13. Cryo-Transmission Electron Microscope 44
3-3. Result and discussion 45
3-3-1. Biocompatible and Self-detachable Photopolymer 45
3-3-2. Biocompatibility and Mechanical Properties. 51
3-3-3. Self-detachable for Open-Access Organ on a Chip 58
3-4. Conclusion 69
Chapter 4. Vascularized tissue transplantation from open-access microfluidic device for regenerative therapy of peripheral artery disease (PAD) 70
4-1. Introduction 70
4-2. Materials and Method 72
4-2-1. 3D Printed Microfluidic Device 72
4-2-2. Photoresin Preparation 74
4-2-3. Surface Treatment 74
4-2-4. Through-hole Membrane Integration 75
4-2-5. Cell Culture and Vasculogenesis Cell Seeding 76
4-2-6. Immunostaining and Imaging 79
4-2-7. Hindlimb Ischemia Model and in vivo Vascularized Hydrogel Implantation 80
4-2-8. Histological analysis and immunostaining 81
4-2-9. Self-detachable for Advanced Analysis of Organ on a Chip 82
4-3. Result and discussion 85
4-3-1. Vascularized Tissue Process and Self-detached Through-hole Membrane 85
4-3-2. Vascularized Tissue Analysis 88
4-3-3. Vascularized Tissue Improves Recovery of Blood Perfusion and Limb salvage in an Ischemic Hind Limb 90
4-4. Conclusion 95
Chapter 5. Concluding Remarks 97
5-1. Conclusion 97
Bibiliography 99
국문초록 112
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dc.formatapplication/pdf-
dc.format.extent10270203 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectOrgan on a Chip-
dc.subjectRegenerative Medicine-
dc.subjectTissue Engineering-
dc.subjectTransplantation-
dc.subjectMicrofluidic-
dc.subject.ddc621-
dc.titleEngineering and Validation of Open Microfluidic Platform for Organ on a Chip-
dc.typeThesis-
dc.description.degreeDoctor-
dc.contributor.affiliation공과대학 기계항공공학부-
dc.date.awarded2018-02-
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College of Engineering/Engineering Practice School (공과대학/대학원)Dept. of Mechanical Aerospace Engineering (기계항공공학부)Theses (Ph.D. / Sc.D._기계항공공학부)
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