Vantyghem M C, Kerr-Conte J, Pattou F, Gevaert M H, Hober C, Defossez A, Mazzuca M, Beauvillain J C
Unité de Soins Normalisée "A", Lille Cedex, France.
Histochem Cell Biol. 1996 Nov;106(5):511-9. doi: 10.1007/BF02473314.
Treatment of diabetes mellitus by transplantation of isolated pancreatic islets could constitute an alternative to human pancreas allograft. Before transplantation, porcine islets are submitted to a procedure of isolation and purification. The quality of islets through these different steps may be assessed by morphological and functional studies. The aim of this work was the histological characterization of the four main cell types of porcine adult endocrine islets during the different steps of the isolation procedure using immunohistochemistry (IHC) applied in light (LM) and electron microscopy (EM). In fresh pancreas, islets were various sizes and shapes in LM. The number was not found different between the different portions of the pancreas. In IHC, insulin (Ins)-secreting cells accounted for the majority of the islet cells, while glucagon(Glu)-somatostatin (Som)- and polypeptide(PP)-immunoreactive cells, in decreasing number, were found in the mantle around the core of Ins-cells. In EM, B-cells contained poly-hedric granules with a dense central core and clear halo. Glu granules were spherical and very dense. D-cells and PP-cells were characterized by numerous granules, rather spherical and of inequal density for Som and more ellipsoidal for PP granules. After purification in Euroficoll, in EM, the four cellular types remained recognizable, but underwent vacuolization, mitochondrial swelling, and enlargement of intercellular spaces. After 3 days of culture on plastic dishes, as on Biopore membranes in a Millicell insert, microvilli appeared and vacuolization increased in EM. At the seventh day of culture, in EM, most of the cells were lysed in contrast to LM where at the same time, the four cell types were clearly identified by IHC but only in collagen matrix. Important discrepancies were noticed between LM and EM. This fact emphasizes the complementarity of morphological and functional studies in assessment of the quality of an islet isolation.
通过移植分离的胰岛治疗糖尿病可能成为人类胰腺同种异体移植的一种替代方法。移植前，猪胰岛需经过分离和纯化程序。可通过形态学和功能研究评估胰岛在这些不同步骤后的质量。本研究的目的是在分离程序的不同步骤中，使用光镜（LM）和电镜（EM）免疫组织化学（IHC）方法对成年猪内分泌胰岛的四种主要细胞类型进行组织学特征分析。在新鲜胰腺中，光镜下胰岛大小和形状各异。胰腺不同部位的胰岛数量未见差异。免疫组织化学显示，分泌胰岛素（Ins）的细胞占胰岛细胞的大多数，而分泌胰高血糖素（Glu）、生长抑素（Som）和多肽（PP）的免疫反应性细胞数量逐渐减少，位于Ins细胞核心周围的外层。电镜下，B细胞含有多面体颗粒，中央核心致密，周围有清晰晕圈。Glu颗粒呈球形且非常致密。D细胞和PP细胞的特征是有许多颗粒，D细胞的颗粒较球形，Som颗粒密度不均，PP颗粒更呈椭圆形。在Euroficoll中纯化后，电镜下四种细胞类型仍可辨认，但出现了空泡化、线粒体肿胀和细胞间隙增大。在塑料培养皿上培养3天后，以及在Millicell插入物中的Biopore膜上培养时，电镜下可见微绒毛出现且空泡化增加。培养第7天，电镜下大多数细胞溶解，而光镜下此时通过免疫组织化学可清楚识别四种细胞类型，但仅在胶原基质中。光镜和电镜之间存在重要差异。这一事实强调了形态学和功能研究在评估胰岛分离质量方面的互补性。
