中华内分泌外科杂志
中華內分泌外科雜誌
중화내분비외과잡지
CHINESE JOURNAL OF ENDOCRINE SURGERY
2011年
5期
299-301
,共3页
冯金发%许评%宋纯%周毅
馮金髮%許評%宋純%週毅
풍금발%허평%송순%주의
胰岛移植%分离%纯化%胰岛细胞
胰島移植%分離%純化%胰島細胞
이도이식%분리%순화%이도세포
Islet transplantation%Isolation%Purification%Islet
目的 探讨影响胰岛分离与纯化结果的相关因素,寻找获得足够数量、高纯度、具有活性的胰岛细胞的方法,为临床胰岛移植应用提供实验基础.方法 寻找自动分离技术连续分离22只犬的胰岛,连续性密度梯度离心法纯化胰岛,观察分离纯化出来的胰岛细胞大体形态、超微结构,用葡萄糖刺激释放实验检验其活性.结果 纯化前胰岛计数平均为( 155 040±310) IEQ/胰腺,纯化后的胰岛平均计数为(74200±185)IEQ/胰腺,纯度约为(89.4±2.6)％,回收率约为(47.8±1.3)％,活率达到(93.0±1.7)％.胰岛分离纯化的结果无论在数量上还是在质量上都随着分离技术的不断更新,操作技术的娴熟而有很大提高.葡萄糖刺激释放实验显示低糖组与高糖组比较P ＜0.001,提示分离纯化后获得的胰岛功能状态良好.结论 从犬中分离和纯化出足够数量的胰岛细胞,可用于临床研究.
目的 探討影響胰島分離與純化結果的相關因素,尋找穫得足夠數量、高純度、具有活性的胰島細胞的方法,為臨床胰島移植應用提供實驗基礎.方法 尋找自動分離技術連續分離22隻犬的胰島,連續性密度梯度離心法純化胰島,觀察分離純化齣來的胰島細胞大體形態、超微結構,用葡萄糖刺激釋放實驗檢驗其活性.結果 純化前胰島計數平均為( 155 040±310) IEQ/胰腺,純化後的胰島平均計數為(74200±185)IEQ/胰腺,純度約為(89.4±2.6)％,迴收率約為(47.8±1.3)％,活率達到(93.0±1.7)％.胰島分離純化的結果無論在數量上還是在質量上都隨著分離技術的不斷更新,操作技術的嫻熟而有很大提高.葡萄糖刺激釋放實驗顯示低糖組與高糖組比較P ＜0.001,提示分離純化後穫得的胰島功能狀態良好.結論 從犬中分離和純化齣足夠數量的胰島細胞,可用于臨床研究.
목적 탐토영향이도분리여순화결과적상관인소,심조획득족구수량、고순도、구유활성적이도세포적방법,위림상이도이식응용제공실험기출.방법 심조자동분리기술련속분리22지견적이도,련속성밀도제도리심법순화이도,관찰분리순화출래적이도세포대체형태、초미결구,용포도당자격석방실험검험기활성.결과 순화전이도계수평균위( 155 040±310) IEQ/이선,순화후적이도평균계수위(74200±185)IEQ/이선,순도약위(89.4±2.6)％,회수솔약위(47.8±1.3)％,활솔체도(93.0±1.7)％.이도분리순화적결과무론재수량상환시재질량상도수착분리기술적불단경신,조작기술적한숙이유흔대제고.포도당자격석방실험현시저당조여고당조비교P ＜0.001,제시분리순화후획득적이도공능상태량호.결론 종견중분리화순화출족구수량적이도세포,가용우림상연구.
Objective To investigate factors influencing islet isolation and purification in dogs and to explore how to obtain massive purified viable islets from canine pancreas.To provide experimental evidence for pancreatic islet transplantation in clinical practice.Methods Islets were isolated from pancreas of 22 dogs by modified automated techniques,followed by purification using continuous density gradients.The purity of islets was determined by dithizone (DTZ) staining,and the function of islets was evaluated by glucose stimulating insulin release test.Results The crude islets yield was ( 155 040 ±310) IEQ/pancreas while the yield was (74 200 ±185) IEQ/pancreas after purification.The average purity was (89.4 ±2.6) ％.The recovery rate was(47.8 ±1.3) ％.Islet viability was(93.0 ± 1.7)％.The modified techniques improved both the yield and the quality in islet isolation and purification.The islets were morphologically intact after purification.Glucose stimulating insulin release test showed that the difference of the secreted insulin concentration had statistical significance ( P ＜0.001 ) between low concentration glucose group and high concentration glucose group.Conclusion It is of great value for clinical research to obtain enough islets through islet isolation and purification in dogs.
