Vajkoczy P, Menger M D
Department of Neurosurgery, Klinikum Mannheim, University of Heidelberg, Mannheim Germany.
Exp Clin Endocrinol Diabetes. 1997;105(3):152-5. doi: 10.1055/s-0029-1211744.
Addition of 10% albumin to the digestion medium has been suggested to enhance yield and integrity of harvested islets by inhibition of proteolytic activities and to improve endocrine function early after transplantation. The aim of this study was to evaluate in vivo by means of intravital fluorescence microscopy whether this rapid reversal of hyperglycemia after transplantation is due to improved graft vascularization. Pancreatic islets were isolated from Syrian golden hamsters by collagenase digestion using either solely Hank's balanced salt solution (HBSS) or HBSS supplemented with 10% human serum albumin. Islets were then transplanted into the dorsal skinfold chamber of syngeneic animals (control: N = 8 animals, n = 50 islets; albumin: N = 7, n = 41). The grafts' microvasculature was analysed on days 6, 10, and 14 after transplantation. Immunohistochemical staining for insulin was performed at the end of the microscopic observation period. Islet isolation with albumin supplementation did not increase islet yield. However, photomicroscopic analysis suggested a beneficial effect on the isolation process with improved islet integrity and prevention of outer margin irregularities, in particular in large islets. Analysis of revascularization 6 days after transplantation revealed in the control group a functional capillary density (FCD) of 477 +/- 47 cm-1. On day 10 FCD increased to 680 +/- 42 cm-1 with no further changes on day 14, indicating complete revascularization. Islets in the albumin group demonstrated a comparable FCD of 598 +/- 49 cm-1 on day 10 and complete revascularization on day 14 (655 +/- 45 cm-1). The angio-architecture of the islets was found similar in both groups, presenting with a glomerulum-like capillary network, comparable to that of pancreatic islets in situ. We conclude that the addition of 10% serum albumin to the collagenase digestion medium improves the preservation of the structural integrity of isolated pancreatic islets, however, does not influence the process of graft vascularization. Thus, improved early graft function may rather be due to superior preservation of islet cell integrity and function.
有人建议在消化培养基中添加10%的白蛋白,以通过抑制蛋白水解活性来提高收获的胰岛的产量和完整性,并改善移植后早期的内分泌功能。本研究的目的是通过活体荧光显微镜在体内评估移植后高血糖的这种快速逆转是否归因于移植组织血管化的改善。使用单纯的汉克平衡盐溶液(HBSS)或补充有10%人血清白蛋白的HBSS,通过胶原酶消化从叙利亚金仓鼠中分离胰岛。然后将胰岛移植到同基因动物的背部皮褶腔中(对照组:N = 8只动物,n = 50个胰岛;白蛋白组:N = 7只,n = 41个)。在移植后第6、10和14天分析移植组织的微血管系统。在显微镜观察期结束时进行胰岛素免疫组织化学染色。添加白蛋白进行胰岛分离并没有增加胰岛产量。然而,显微分析表明对分离过程有有益影响,可改善胰岛完整性并防止外缘不规则,特别是在大胰岛中。移植后6天对血管再生的分析显示,对照组的功能毛细血管密度(FCD)为477±47 cm-1。在第10天FCD增加到680±42 cm-1,在第14天没有进一步变化,表明血管再生完成。白蛋白组的胰岛在第10天显示出相当的FCD为598±49 cm-1,在第14天血管再生完成(655±45 cm-1)。发现两组胰岛的血管结构相似,呈现肾小球样毛细血管网络,与原位胰腺胰岛的网络相当。我们得出结论,在胶原酶消化培养基中添加10%血清白蛋白可改善分离的胰腺胰岛结构完整性的保存,但不影响移植组织的血管化过程。因此,移植组织早期功能的改善可能更多地归因于胰岛细胞完整性和功能的更好保存。