中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2007年
38期
7701-7705
,共5页
高宏君%梁泰生%杨欢%罗向东%吴佩钟%谭臻%梁芳芳
高宏君%樑泰生%楊歡%囉嚮東%吳珮鐘%譚臻%樑芳芳
고굉군%량태생%양환%라향동%오패종%담진%량방방
胰岛细胞%冷缺血时间%组织相容性%移植
胰島細胞%冷缺血時間%組織相容性%移植
이도세포%랭결혈시간%조직상용성%이식
背景:在胰岛移植过程中,胰岛细胞的数量和胰岛细胞的活性是非常重要的.胰岛细胞移植中玲缺血时间和组织相容性是关系胰岛细胞移植能否成功的关键问题.目的:观察冷缺血时间及血液相容性不同对于胰岛细胞数量和活性的影响.设计:观察性实验.单位:广西中医学院附属瑞康医院.材料:采用脑死亡自愿捐赠者的器官(已知血型和HLA配型),在其他器官切取后或同时进行胰腺的获取;血型相同、HLA配型相符的血液;HLA配型高致敏(错配3个以上位点)或群体反应性抗体＞50%或淋巴毒阳性受者的血液;分离纯化后的胰岛细胞悬浊液,经过70μm的筛网滤过后,保留直径大于70μm的胰岛细胞,配置成浓度1.20×105L-1左右活性胰岛细胞悬浊液.方法:高渗枸橼酸盐嘌呤溶液经过腹主动脉原位灌注后,参照文献进行肝、肾、胰腺联合及分别切取.用于切取的时间:胰腺肾脏的联合切取、肝脏胰腺肾脏的联合切取方法平均需时15 min,肝脏、胰腺、肾脏的单独切取平均需时20 min,用于胰岛分离的胰腺12份,冷缺血时间2.5～8 h.冷却血时间在5 h以内的9份,温缺血时间0～3 min,消化的时间15±2.4 min.采用胶原霉P进行消化分离胰岛细胞,双硫腙、丫腚橙染色测定不同的冷缺血时间条件下活性胰岛细胞活性,分析体外胰岛细胞和血液与供受者的组织相容性和胰岛细胞存活的关系.主要观察指标:胰岛细胞活性率和血小板、中性粒细胞和单核细胞计数.结果:肝、肾、胰腺联合切取及各器官的单独切取顺利,在玲却血5 h以内胰岛细胞活性率都在80%以上,用于胰岛细胞移植的胰腺和其他器官的切取不会影响胰岛细胞的活性;人类胰岛暴露于未经抗凝的人类血中,胰岛将诱发一个迅速的血细胞消耗;进行血小板、中性粒细胞和单核细胞计数,HLA错配组和HLA匹配组与对照组比较血细胞都发生明显的消耗;加入肝素后血细胞计数与对照组比较差异明显(P＜0.05),反应明显减轻.HLA匹配组和HLA错配组胰岛细胞体外培养24 h活性胰岛细胞数量较,差异明显(P＜0.05).结论:在冷缺血时间＜5 h的情况下获取的胰腺可以用于临床胰岛细胞的移植;血液相容性好能够明显提高胰岛细胞移植的成功率.
揹景:在胰島移植過程中,胰島細胞的數量和胰島細胞的活性是非常重要的.胰島細胞移植中玲缺血時間和組織相容性是關繫胰島細胞移植能否成功的關鍵問題.目的:觀察冷缺血時間及血液相容性不同對于胰島細胞數量和活性的影響.設計:觀察性實驗.單位:廣西中醫學院附屬瑞康醫院.材料:採用腦死亡自願捐贈者的器官(已知血型和HLA配型),在其他器官切取後或同時進行胰腺的穫取;血型相同、HLA配型相符的血液;HLA配型高緻敏(錯配3箇以上位點)或群體反應性抗體＞50%或淋巴毒暘性受者的血液;分離純化後的胰島細胞懸濁液,經過70μm的篩網濾過後,保留直徑大于70μm的胰島細胞,配置成濃度1.20×105L-1左右活性胰島細胞懸濁液.方法:高滲枸櫞痠鹽嘌呤溶液經過腹主動脈原位灌註後,參照文獻進行肝、腎、胰腺聯閤及分彆切取.用于切取的時間:胰腺腎髒的聯閤切取、肝髒胰腺腎髒的聯閤切取方法平均需時15 min,肝髒、胰腺、腎髒的單獨切取平均需時20 min,用于胰島分離的胰腺12份,冷缺血時間2.5～8 h.冷卻血時間在5 h以內的9份,溫缺血時間0～3 min,消化的時間15±2.4 min.採用膠原黴P進行消化分離胰島細胞,雙硫腙、丫腚橙染色測定不同的冷缺血時間條件下活性胰島細胞活性,分析體外胰島細胞和血液與供受者的組織相容性和胰島細胞存活的關繫.主要觀察指標:胰島細胞活性率和血小闆、中性粒細胞和單覈細胞計數.結果:肝、腎、胰腺聯閤切取及各器官的單獨切取順利,在玲卻血5 h以內胰島細胞活性率都在80%以上,用于胰島細胞移植的胰腺和其他器官的切取不會影響胰島細胞的活性;人類胰島暴露于未經抗凝的人類血中,胰島將誘髮一箇迅速的血細胞消耗;進行血小闆、中性粒細胞和單覈細胞計數,HLA錯配組和HLA匹配組與對照組比較血細胞都髮生明顯的消耗;加入肝素後血細胞計數與對照組比較差異明顯(P＜0.05),反應明顯減輕.HLA匹配組和HLA錯配組胰島細胞體外培養24 h活性胰島細胞數量較,差異明顯(P＜0.05).結論:在冷缺血時間＜5 h的情況下穫取的胰腺可以用于臨床胰島細胞的移植;血液相容性好能夠明顯提高胰島細胞移植的成功率.
배경:재이도이식과정중,이도세포적수량화이도세포적활성시비상중요적.이도세포이식중령결혈시간화조직상용성시관계이도세포이식능부성공적관건문제.목적:관찰랭결혈시간급혈액상용성불동대우이도세포수량화활성적영향.설계:관찰성실험.단위:엄서중의학원부속서강의원.재료:채용뇌사망자원연증자적기관(이지혈형화HLA배형),재기타기관절취후혹동시진행이선적획취;혈형상동、HLA배형상부적혈액;HLA배형고치민(착배3개이상위점)혹군체반응성항체＞50%혹림파독양성수자적혈액;분리순화후적이도세포현탁액,경과70μm적사망려과후,보류직경대우70μm적이도세포,배치성농도1.20×105L-1좌우활성이도세포현탁액.방법:고삼구연산염표령용액경과복주동맥원위관주후,삼조문헌진행간、신、이선연합급분별절취.용우절취적시간:이선신장적연합절취、간장이선신장적연합절취방법평균수시15 min,간장、이선、신장적단독절취평균수시20 min,용우이도분리적이선12빈,랭결혈시간2.5～8 h.냉각혈시간재5 h이내적9빈,온결혈시간0～3 min,소화적시간15±2.4 min.채용효원매P진행소화분리이도세포,쌍류종、아정등염색측정불동적랭결혈시간조건하활성이도세포활성,분석체외이도세포화혈액여공수자적조직상용성화이도세포존활적관계.주요관찰지표:이도세포활성솔화혈소판、중성립세포화단핵세포계수.결과:간、신、이선연합절취급각기관적단독절취순리,재령각혈5 h이내이도세포활성솔도재80%이상,용우이도세포이식적이선화기타기관적절취불회영향이도세포적활성;인류이도폭로우미경항응적인류혈중,이도장유발일개신속적혈세포소모;진행혈소판、중성립세포화단핵세포계수,HLA착배조화HLA필배조여대조조비교혈세포도발생명현적소모;가입간소후혈세포계수여대조조비교차이명현(P＜0.05),반응명현감경.HLA필배조화HLA착배조이도세포체외배양24 h활성이도세포수량교,차이명현(P＜0.05).결론:재랭결혈시간＜5 h적정황하획취적이선가이용우림상이도세포적이식;혈액상용성호능구명현제고이도세포이식적성공솔.
BACKGROUND: The quantity and bioactivity of isolated islet are vital to islet transplantation; while, the cold ischemia time and human leucocyte antigen (HLA) typing are key factors to islet quantity and bioactivity which inflect islet transplantation.OBJECTIVE: To observe the effects of cold ischemia time and blood compatibility on quantity and bioactivity of islet cells.DESIGN: Observational study.SETTING: Ruikang Affiliated Hospital of Guangxi College of Traditional Chinese Medicine.MATERIALS: Organs from voluntary donors died of irreversible coma were adopted whose blood-type and HLA typing had been known, pancreases acquisition was carried on after other organs ablation or in the meantime. The blood of identical ABO and HLA matching conformity, or HLA cross-matching hypersensitization (missmatching over 3 Iocuses),or panel reaction antibody (PRA) ＞ 50%, or lymphocyte cytotoxin crossmatching test positive. The isolated and purified islet suspension was filtered by 70 μm filter, which result in preparing 1.2×105/L islet suspention.METHODS: Hypertonic citrate adenine solution was perfused into aorta, and kidney-pancreases and kidney-pancreases-liver were cut together or kidney-pancreases-liver was cut separately. Islet activity was judged by diphenylthiocarbazone (DTZ) dyeing and acridine orange (AO) dyeing; meanwhile, twelve pancreases far from contamination were aquired, mean ablation time was 15 minutes; cold ischemia time ranged from 2.5 to 8 hours. Cold ischemia time of nine pancreases was controlled in 5 hours, warm ischemia time was 0-3 minutes, and peptic time was (15±2.4) minutes, correlation of islet cells and histocompatibility with survival of islet cells was analyzed.MAIN OUTCOME MEASURES: Survival rate of islet cells; counts of platelet, heterophil granulocyte and monocyte.RESULTS: The cutting of kidney, pancreases and liver were successful. If the cold ischemia time was controlled within 5 hours, activity of islets was above 80%. Pancreatic gland used for islet transplantation and cutting of other organs could not affect activity of islet cells. When human islets were exposed to human blood, it would induce a rapid consumption of platelets, neutrophils, monocytes and lymphocytes in the blood. Consumption of blood cells was more in the HLA typing groups than that in the control group. After adding heparin, there was significant difference in cell account among the three groups (P ＜ 0.05) and the reaction was relieved obviously. After 24-hour cultivation, there were significant difference in active islets quantity between HLA typing compatibility group and HLA typing incompatibility group (P ＜ 0.05).CONCLUSION: Pancreatic gland obtained under the cold ischemia time ＜ 5 hours can be used in clinical transplantation of islet cells; a good histocompatibility can raise successful rate of transplantation of islet cells.
