Min T, Yi L, Chao Z, Haitao Z, Wei W, Liang Y, Bo W
Department of Hepatobiliary Surgery, First Affiliated Hospital, Medical College, Xi'an Jiaotong University, Xi'an, People's Republic of China.
Transplant Proc. 2010 Jun;42(5):1825-9. doi: 10.1016/j.transproceed.2010.01.068.
Sufficient and favorable biological functions of islets are major problems hindering xenotransplantation. The aim of the present study was to evaluate the effects on harvesting, purity, viability, and function of using improved Visipaque (iodixanol) and Ficoll-400 for adult porcine islet purification.
Twelve adult porcine pancreata were randomly divided into an Iodixanol-University of Wisconsin (UW) group and a Ficoll-400-UW group according to the purification method. Porcine pancreata were isolated by collagenase digestion. After isolation and purification, the islet yield and purity were evaluated by dithizone staining, and islet function assessed by in vitro insulin release assays and in vivo islet xenotransplantation.
There were no marked differences in the islet yield before purification (5254.67 +/- 189.44 IEQ/g vs 5092.67 +/- 178.94 IEQ/g, P > .05). After purification, there were significantly more islets harvested in Iodixanol-UW group than in the Ficoll-400-UW group: 4222.00 +/- 228.84 IEQ/g vs 3036.83 +/- 79.60 IEQ/g (P < .05). Islets from the two groups showed satisfactory insulin secretory ability. There were no significant differences in islet survival times between the two groups in diabetic rats: 8.2 +/- 1.619 days vs 6.9 +/- 1.197 days (P > .05).
The improved iodixanol-UW density gradient method was superior to Ficoll-400 method to improve the number, viability, and insulin secret of purified adult porcine islets although the benefits did not improve in vivo survival.
胰岛充足且良好的生物学功能是阻碍异种移植的主要问题。本研究的目的是评估使用改良的威视派克(碘克沙醇)和聚蔗糖-400进行成年猪胰岛纯化对收获量、纯度、活力和功能的影响。
根据纯化方法将12个成年猪胰腺随机分为碘克沙醇-威斯康星大学(UW)组和聚蔗糖-400-UW组。通过胶原酶消化分离猪胰腺。分离和纯化后,用双硫腙染色评估胰岛产量和纯度,通过体外胰岛素释放试验和体内胰岛异种移植评估胰岛功能。
纯化前的胰岛产量无显著差异(5254.67±189.44 IEQ/g对5092.67±178.94 IEQ/g,P>.05)。纯化后,碘克沙醇-UW组收获的胰岛明显多于聚蔗糖-400-UW组:4222.00±228.84 IEQ/g对3036.83±79.60 IEQ/g(P<.05)。两组的胰岛均显示出令人满意的胰岛素分泌能力。糖尿病大鼠两组间的胰岛存活时间无显著差异:8.2±1.619天对6.9±1.197天(P>.05)。
改良的碘克沙醇-UW密度梯度法在提高纯化的成年猪胰岛数量、活力和胰岛素分泌方面优于聚蔗糖-400法,尽管其益处并未改善体内存活情况。