摘要
目的对国际用于人胰岛制备的两种胶原酶(Serva NB1胶原酶和VitacyteGOLD胶原酶)在中国供者胰岛分离中的效果进行比较。探讨适合中国人的胰岛分离方法。方法入选17例供者胰腺均来自死亡后器官捐献,其中12例使用Serva NB1胶原酶消化,5例使用Vitacyte GOLD胶原酶消化。在百级层流实验室中进行胰腺分离,经过修整,分别应用不同胶原酶充分灌注,利用Ricordi方法消化并记录消化时间,利用连续密度梯度法纯化胰岛,然后检测胰岛产率、活性和葡糖糖刺激的胰岛素分泌能力,以评估分离胰岛的数量和质量。结果Vitacyte GOLD胶原酶比Serva NB1胶原酶所需的消化时间更短(P<0.05)。纯化后直径在50~100 μm之间的胰岛,Vitacyte GOLD胶原酶组明显多于ServaNB1胶原酶组(P<0.05),直径在251~300 μm、301~350 μm之间的胰岛,Vitacyte GOLD胶原酶组明显少于ServaNB1胶原酶组(P<0.05)。两种胶原酶分离获得总胰岛数量差异无统计学意义(P>0.05)。两种胶原酶分离胰岛的活性检测和GSI结果差异无统计学意义(P>0.05)。结论Serva NB1胶原酶和Vitacyte GOLD胶原酶均可用于临床分离制备中国人胰岛。
Objective To retrospectively compare the efficacy of Serva NB1 collagenase with Vitacyte GOLD collagenase on islet isolation of pancreas. MethodsAll the human pancreata were obtained from Chinese organ donors. In GMP laboratory, the pancreata were trimmed and distended with Serva NB1 collagenase (Serva NB1, n=12) or Vitacyte GOLD collagenase (Vitacyte GOLD, n=5) and digested according to a modified Ricordi semi-automatic protocol, and the digestion duration was recorded. The digested islets were then collected and washed, followed by the continuous density purification in a Cobe 2991 cell separator. The islet yield, purity, viability and glucose-stimulated insulin release (GSI) were determined each time after purification. Quantity and quality of isolated islets were determined by digestion efficacy. ResultsThe digestion duration in Vitacyte GOLD collagenase group was significantly shorter than in Serva NB1 collagenase group to achieve the same digestion endpoint (P<0.05). The islets yields of different sizes were variable between the two groups. The Vitacyte GOLD collagenase digestion produced more islets with a diameter range of 50-100 μm than the ServaNB1 collagenase digestion (P<0.05), but the latter yielded more islets with a diameter range of 251-300 μm and 301-350 μm (P<0.05). There was no significant difference in total islets yields, viability, and GSI between two collagenase digestions (P>0.05). ConclusionBoth Vitacyte GOLD collagenase and Serva NB1 collagenase can be used for the clinical islet isolation in China.
作者
刘尧娟
邹家琦
王贯乔
王乐
刘腾丽
王树森
王智平
裴广辉
王金山
沈中阳
Liu Yaojuan;Zou Jiaqi;Wang Guanqiao;Wang Le;Liu Tengli;Wang Shusen;Wang Zhiping;Pei Guanghui;Wang Jinshan;Shen Zhongyang(Organ Transplantation Center,Tianjin First Central Hospital,Tianjin 300192,China)
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2018年第12期720-724,共5页
Chinese Journal of Organ Transplantation
基金
国家重点研发计划(2016YFC1305104)
天津市器官移植临床研究中心(15ZXLCSY00070)
药物化学生物学国家重点实验室开放基金(2018016).
关键词
胰岛移植
胰岛分离
胶原酶
Islet isolation
Collagenase
Islet transplantation
