초록 ▼

연구개발사업의 목표 :
1) 심장혈관계 장기를 냉동보존하여 심장수술에 사용
- 대동맥판막 및 대동맥의 냉동보존, 폐동맥판막 및 폐동맥팻취의 냉동보존, 복재정맥 및 소동맥의 냉동보존
2) 조직처리 중 감염여부 확인 및 조직을 통한 질환전이를 예방하는 검사법 개발

연구방법 :
1) 1차연구에서 개발된 대동맥판막의 냉동보존방법을 적용한 제반 심혈관계 조직의 보존
- 현재 개발된 대동맥판막의 냉동보존법의 재검토
- 심장수술시 남는 복재정맥 및 내흉동맥의 냉동보존시행
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연구개발사업의 목표 :
1) 심장혈관계 장기를 냉동보존하여 심장수술에 사용
- 대동맥판막 및 대동맥의 냉동보존, 폐동맥판막 및 폐동맥팻취의 냉동보존, 복재정맥 및 소동맥의 냉동보존
2) 조직처리 중 감염여부 확인 및 조직을 통한 질환전이를 예방하는 검사법 개발

연구방법 :
1) 1차연구에서 개발된 대동맥판막의 냉동보존방법을 적용한 제반 심혈관계 조직의 보존
- 현재 개발된 대동맥판막의 냉동보존법의 재검토
- 심장수술시 남는 복재정맥 및 내흉동맥의 냉동보존시행
- 국립과학수사연구소와의 공조로 공여받을 수 있는 장기를 이용한 냉동보존법의 검증
2) 조직처리 중 감염여부 확인 및 조직을 통한 질환전이를 예방하는 검사법 개발
- 인간에게 이식가능한 감염, 암, 전신질환의 전이를 방지할 수 있는 검사법 개발
- 조직의 적출, 이송, 보존처리, 보관, 재이송 전반의 과정에서 감염의 가능성을 줄이는 동시에 감염여부를 확인하는 체제를 개발
- 위의 방법론을 체계화 하여 기타 조직은행의 감염검사의 표준을 제시

Abstract ▼

The Purpose of Project
1) development of cardiopvascular tissues by cryopreservation for cardiovascular surgerydevelopment of cryopreservation method of aortic valve, aorta, pulmonic valve, pulmonic patch, pericardium, saphenous vein andfemoral artery
2) development of infection and disease

The Purpose of Project
1) development of cardiopvascular tissues by cryopreservation for cardiovascular surgerydevelopment of cryopreservation method of aortic valve, aorta, pulmonic valve, pulmonic patch, pericardium, saphenous vein andfemoral artery
2) development of infection and disease transmission control using the test for infection during harvesting, processing and storage

Method and Results
1) cryopreservation of cardiovascular tissues was developed by previously confirmed cryopreservation method of aortic valve
Review of previously confirmed cryopreservation method. Cryopreservation of remnat of saphenous vein and internal thoracic arteryafter coronary bypass surgery. Development of cryopreservation methods for canine carotid artery and jugular vein.
Searching method of the ideal freezing curve was developed by indirect calculation method and it was confirmed as useful tool for various cardiovascular tissue preservation. The efficacy was evaluated by cell viability test after preservation of canine jugular vein with new freezing curve
Cryopreservation method of saphenous vein was developed using stopcock and thread without sten. Two different length packagewas also developed for 15cm and 20cm vein. However, arterial graft was not effectively preserved because of small diameter which couldn't contain cryoprotectant. This method was evaluated by canine carotid artery implantation experiments.
For quality control of tissue processing, we developed complete set of document forms and procedural manual. These forms were filled up using web-based database program. Yonsei cardiovascular tissue bank was opened and 30 researchers attendedworkshop for studying how to harvest, process, freeze and thaw. This document and manual will be the standard for cardiovascular tissue processing in my country. My tissue bank already have 11 homograft and release 14 homograft which were successfully implanted. And also 2 homograft were released to other hospital.
The animal experiments of canine carotid artery implantation using cryopreserved carotid artery and jugular vein were performed. In the first year, we performed implantation of cryopreserved artery, sham artery and fresh allograft artery in one side and cryopreserved vein, sham vein and fresh allograft vein in other side. The implantation reaction of cryopreserved artery and vein was not good as freshartery and vein. In the second year.
We performed implantation of cryopreserved artery and vein comparing implantation of freshartery and vein. The result was similar with the result of the first year, and we can think that result was not excellent however, cryopreserved artery and vein will be useful conduit for redo coronary artery surgery or peripheral vascular surgery because of long term storage and large numbers

2) In terms of impacts on cell viability by pre-treatment with antibiotic solution, it was considered that the decisive factor decreasingthe viability of Endothelial cells (EC) as well as whole cells (WC) might be the duration of retention in an antibiotic solutionin the short
-term cryopreservation of cardiovascular organs

Three different methods- trypan blue staining assay, [$^3H$]-proline incorporation assay, and flow cytometry (FCM) combined with fluorescein is othiocyanate (FICT) conjugated with GSA (Griffonia simplicifolia agglutin) (GSA-FITC)/propidium iodide (PI) double staining - were compared to determine the most efficient evaluation method for the viability of EC viability dissociated from venous tissue. GSA-FITC/PI double staining FCM analytical method could rapidly obtain the viability of EC and WC from the same tissue at once without separating EC from WC, and also obtain viability of the cells in each step of cryopreservation process.

목차 Contents

• I. 연구개발결과 요약문...4
• (한글) 냉동보존된 심혈관계 장기의 개발...4
• (영문) Development of Cryopreserved Cardiovascular Tissues...6
• Ⅱ. 총괄연구개발과제 연구결과...8
• 1. 총괄연구개발과제의 최종 연구개발 목표...8
• 2. 총괄연구개발과제의 최종 연구개발 내용 및 결과...26
• 3. 총괄연구개발과제의 연구결과 고찰 및 결론...153
• 4. 총괄연구개발과제의 연구성과 및 목표달성도...155
• 5. 총괄연구개발과제의 활용계획...164
• 6. 첨부서류...168
• Ⅲ. 제 1세부연구개발과제 연구결과 (세부과제별로 작성)...187
• 1. 제 1세부연구개발과제의 최종 연구개발 목표...188
• 2. 제 1세부연구개발과제의 연구대상 및 방법...192
• 3. 제 1세부연구개발과제의 최종 연구개발결과...198
• 4. 제 1세부연구개발과제의 연구결과 고찰 및 결론...312
• 5. 제 1세부연구개발과제의 연구성과 및 목표달성도...313
• 6. 제 1세부연구개발과제의 활용계획...314
• 7. 참고문헌...317
• Ⅳ. 제 2세부연구개발과제 연구결과 (세부과제별로 작성)...319
• 1. 제 2세부연구개발과제의 최종 연구개발 목표...320
• 2. 제 2세부연구개발과제의 연구대상 및 방법...323
• 3. 제 2세부연구개발과제의 최종 연구개발결과...333
• 4. 제 2세부연구개발과제의 연구결과 고찰 및 결론...382
• 5. 제 2세부연구개발과제의 연구성과 및 목표달성도...383
• 6. 제 2세부연구개발과제의 활용계획...384
• 7. 참고문헌...384
• 부록...387