Huh K H, Lee J-I, Kim J Y, Jeong J H, Fang Y, Park Y-J, Kang C M, Kim Y S
Research Institute for Transplantation, Yonsei University College of Medicine, Seoul, Korea.
Transplant Proc. 2009 Jan-Feb;41(1):323-5. doi: 10.1016/j.transproceed.2008.10.077.
Porcine-specific obstacles in islet isolation frequently result from the low purity or contamination with exocrine tissues. We implemented a new technique involving as capsulation of islets with excess exocrine tissue as a beneficial material to address those difficulties. Pig islets were hand-picked as high purity (HI) or low purity (LO) islets containing significant amounts of exocrine tissue. We performed static (ST) or shaking (SK) cultures of HI and LO islets. Islet function was examined after 24 hours by a glucose challenge test. Insulin secretion into the culture media was continuously measured using ELISA during a 6-day culture period. Islet function after 24 hours exhibited better maintenance under SK than ST culture as assessed by a stimulation index. The ideal islet morphology was seen in LO islets at 3 days of SK culture with typical islet shapes of a smooth surface and a spherical configuration. In contrast, typical islet morphology was not observed in HI islets under SK culture; maintenance of typical spherical appearances was difficult. Insulin secretion from LO islets under SK culture was higher than under other conditions during the 6-day period. Under SK culture conditions, exocrine-encapsulated LO islets showed enhanced islet function by condensing loose islet aggregates into firm spheroids with native exocrine tissues as a natural scaffold.
胰岛分离过程中猪特异性的障碍常常源于纯度低或受到外分泌组织污染。我们实施了一项新技术,即将含有过量外分泌组织的胰岛进行包封,将外分泌组织作为一种有益材料来解决这些难题。猪胰岛被挑选为含有大量外分泌组织的高纯度(HI)或低纯度(LO)胰岛。我们对HI和LO胰岛进行了静态(ST)或振荡(SK)培养。在24小时后通过葡萄糖刺激试验检测胰岛功能。在为期6天的培养期内,使用酶联免疫吸附测定法(ELISA)持续测量培养基中的胰岛素分泌量。通过刺激指数评估,24小时后的胰岛功能在SK培养条件下比ST培养条件下维持得更好。在SK培养3天时,LO胰岛呈现出理想的胰岛形态,表面光滑,呈典型的球形。相比之下,在SK培养条件下HI胰岛未观察到典型的胰岛形态;难以维持典型的球形外观。在6天培养期内,SK培养条件下LO胰岛的胰岛素分泌高于其他条件下。在SK培养条件下,外分泌组织包封的LO胰岛通过将松散的胰岛聚集体凝聚成坚实的球体,以外分泌组织作为天然支架,从而增强了胰岛功能。