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광범위 창상 치유를 위한 인체 상피세포의 시험관내 배양에 관한 연구

Other Titles
 (A) study on in vitro cultivation of human epithelial cells for covering extensive wound 
Issue Date
1983
Description
의학과/석사
Abstract
[한글] 화상으로 인한 광범위 심층 피부결손은 체내의 수분, 전해질 및 단백질의 손실과 손상된 피부를 복구하기 위해 소요되는 오랜 기간동안 동반되는 폐혈증, 신 부전증, 호흡기 계통의 합병증 등의 치명적인 합병증의 병발로 인한 높은 사망률을 저하시키기 위해 빠른 시일내에 복구되어야 한다. 광범위 화상의 경우 광여피부의 부족으로 전통적인 식피술로는 이를 해결하기 어려운 상태이다. 이글 극복하기 위해 환자의 피부로부터 적은 량의 피부를 조직배양 및 세포배양으로 팽창시켜 다시 재이식하려는 연구가 활발히 진행되고 있는 실정이다. 본 실험에서는 인체의 피부조직으로부터 채취한 상피세포를 시험관 내에서 약 50배로 팽창, 증식시켜 다음과 같은 결과를 얻었다. 1. 시험관 내에서 배양된 상피세포의 증식상태 채취후 1㎣이하의 작은 조각으로 나뉘어진 피부 조직편은 분주후 약 48시간 내지 72시간내에 배양용기 및 지지표면에 부착되 out growth 되었다. 약 2주까지 증식속도가 빨랐으나 그후 3주까지는 증식의 정도가 완만하였 다. 분주후 약 7∼8일 경부터 정상적인 상태의 피세포외에 색소를 갖는 비교적 큰 세포가 조직편으로부터 방사상으로 퍼져 나왔으며 조직편으로부터 증식되어 나오는 상피세포와 조직편분주시 떨어져 나와 독립적으로 증식된 상피세포와 접합되는 양상을 보이면서 증식되었다. 유리된 상피세포는 24시간 내지 48시간 사이에 배양용기 및 지지표면에 부착된 후 48시간 내지 72시간에 집합체를 이루면서 부분적으로 plating되었으며 분주후 4∼5일 사이에 대다수의 상피세포들이 plating되면서 증식의 속도는 가속화되어 약 2주후에는 피부 조직편으로부터 증식된 상피세포와 거의 단일상피 세포층을 형성하였으나 그후 3주까지의 성장속도는 둔화되었다. 2. 상피세포 공여자의 연령에 따른 상피세포의 증식정도 신생아의 foreskin으로부터 얻은 피부 조직편 및 나이가 많은 공여자의 그것보다 분리되는 단일 상피세포의 숫자가 많았으며 세포의 Colony형성과 증식 정도도 빨랐다. 3. 지지표면에 따른 상피세포의 증식정도 유리나 플라스틱보다 Collagen gel위에서 상피세포의 plating효율이 높았으나 증식정도에는 큰 차이를 발견치 못하였다. A study on in vitro cultivation of human epithelial cells for covering extensive wound Hae Kwang Cheong, M.D. Dept. of medical science, The Graduate School, Yonsei University (Directed by Professor Joo Deuk Kim, M.D.) Timely covering for extensive wound is essential for the survival of the patient. Needless to emphasize, autologous skin of the patient is the best of choice for the coverage. However, it is almost impossible to hale enough skin from patient who is suffering from extensive burn. Therefore, various materials , such as, porcine skin, human amniotic membranes, and artificial skin have been used to substitute the autologous skin. Nevertheless, none of the materials truely benefical in prolong the life expectancy of the patients. The use of cultured patient epithelial cert Bs for the source of wound covering and recently this had been tried in clinics and found to be a meat favorable method to solve the fundamental problems involved include difficulties in preparing the cells, funtional defects of the covered epithelium in various aspects. Among those problems, successful explantation of freshly prepared skin into in vitro system would be the major task to be studied. Therefore, in this experiment the general characteristics of freshly explanted human epithelial cells in vitro were studied. The results are as follows ; 1. Epithelial cell multiplication from skin explants and epithelial cells; After cutting skin into fine mince (<1 cu mm) and plating on plastics and collagen gels, the skin explants and epithelial cells settled to the surface and attached to it within 48-72 hours. after attachment, epithelial cells migrated actively and colonies of several cells were gradually formed and epithelial cells appeared at the edge of skin explants. Epithelial cert is multiplication and outgrowth from skin explants appeared actively to about 2 weeks, and it had been grown into mono-epithelial cell layer. Growth rate of epithelial cells decreased after 2 weeks. 2. Difference of growth characteristics between newborn epithelial cell and older donor; Numbers of epithelial cell separated from skin explants during mincing, colony formation efficiency, plating efficiency, and epithelial cell growth rate in newborn was higher than older donor. 3. Difference of growth characteristics between plastic surface and collagen gel ; Colony formation efficiency and plating efficiency of epithelial cells on collagen gel was higher than its on plastic surface. But, no difference in epithelial cell growth rate was found.
[영문] Timely covering for extensive wound is essential for the survival of the patient. Needless to emphasize, autologous skin of the patient is the best of choice for the coverage. However, it is almost impossible to hale enough skin from patient who is suffering from extensive burn. Therefore, various materials , such as, porcine skin, human amniotic membranes, and artificial skin have been used to substitute the autologous skin. Nevertheless, none of the materials truely benefical in prolong the life expectancy of the patients. The use of cultured patient epithelial cert Bs for the source of wound covering and recently this had been tried in clinics and found to be a meat favorable method to solve the fundamental problems involved include difficulties in preparing the cells, funtional defects of the covered epithelium in various aspects. Among those problems, successful explantation of freshly prepared skin into in vitro system would be the major task to be studied. Therefore, in this experiment the general characteristics of freshly explanted human epithelial cells in vitro were studied. The results are as follows ; 1. Epithelial cell multiplication from skin explants and epithelial cells; After cutting skin into fine mince (<1 cu mm) and plating on plastics and collagen gels, the skin explants and epithelial cells settled to the surface and attached to it within 48-72 hours. after attachment, epithelial cells migrated actively and colonies of several cells were gradually formed and epithelial cells appeared at the edge of skin explants. Epithelial cert is multiplication and outgrowth from skin explants appeared actively to about 2 weeks, and it had been grown into mono-epithelial cell layer. Growth rate of epithelial cells decreased after 2 weeks. 2. Difference of growth characteristics between newborn epithelial cell and older donor; Numbers of epithelial cell separated from skin explants during mincing, colony formation efficiency, plating efficiency, and epithelial cell growth rate in newborn was higher than older donor. 3. Difference of growth characteristics between plastic surface and collagen gel; Colony formation efficiency and plating efficiency of epithelial cells on collagen gel was higher than its on plastic surface. But, no difference in epithelial cell growth rate was found.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/116987
Appears in Collections:
2. 학위논문 > 1. College of Medicine (의과대학) > 석사
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