Nacher Montserrat, Estil Les Elisabet, Garcia Ainhoa, Nadal Belen, Pairó Mar, Garcia Cristofer, Secanella Lluís, Novials Anna, Montanya Eduard
Hospital Universitari Bellvitge-IDIBELLL, Hospitalet de Llobregat, Barcelona, Spain.
Cell Transplant. 2016;25(2):343-52. doi: 10.3727/096368915X688119. Epub 2015 May 7.
There is conflicting evidence favoring both the use of human serum (HS) and of human serum albumin (HSA) in human islet culture. We evaluated the effects of HS versus HSA supplementation on 1) in vitro β-cell viability and function and 2) in vivo islet graft revascularization, islet viability, β-cell death, and metabolic outcome after transplantation. Islets isolated from 14 cadaveric organ donors were cultured for 3 days in CMRL 1066 medium supplemented with HS or HSA. After 3 days in culture, β-cell apoptosis was lower in HS group (1.41 ± 0.27 vs. 2.38 ± 0.39%, p = 0.029), and the recovery of islets was 77 ± 11% and 54 ± 1% in HS- and HSA-cultured groups, respectively. Glucose-stimulated insulin secretion (GSIS) was higher in HS group (29.4, range 10.4-99.9, vs. 22.3, range 8.7-70.6, p = 0.031). In vivo viability and revascularization was determined in HS- and HSA-cultured islets transplanted into the anterior chamber of the eye of Balb/c mice (n = 14), and β-cell apoptosis in paraffin-embedded mouse eyes. Islet viability and β-cell apoptosis were similar in both groups. Revascularization was observed in one graft (HS group) on day 10 after transplantation. Islet function was determined in streptozotocin (STZ)-diabetic nude mice (n = 33) transplanted with 2,000 IEQs cultured with HS or HSA that showed similar blood glucose levels and percentage of normoglycemic animals over time. In conclusion, human islets cultured in medium supplemented with HS showed higher survival in vitro, as well as islet viability and function. The higher in vitro survival increased the number of islets available for transplantation. However, the beneficial effect on viability and function did not translate into an improved metabolic evolution when a similar number of HSA- and HS-cultured islets was transplanted.
在人胰岛培养中，支持使用人血清（HS）和人血清白蛋白（HSA）的证据相互矛盾。我们评估了补充HS与HSA对以下方面的影响：1）体外β细胞活力和功能；2）体内胰岛移植后的血管再生、胰岛活力、β细胞死亡及代谢结果。从14名尸体器官捐献者分离的胰岛在补充有HS或HSA的CMRL 1066培养基中培养3天。培养3天后，HS组的β细胞凋亡率较低（1.41±0.27%对2.38±0.39%，p = 0.029），HS培养组和HSA培养组的胰岛回收率分别为77±11%和54±1%。HS组的葡萄糖刺激胰岛素分泌（GSIS）更高（29.4，范围10.4 - 99.9，对22.3，范围8.7 - 70.6，p = 0.031）。将培养有HS和HSA的胰岛移植到Balb/c小鼠（n = 14）眼前房，测定体内活力和血管再生情况，并检测石蜡包埋小鼠眼中的β细胞凋亡。两组的胰岛活力和β细胞凋亡相似。移植后第10天，在一个移植物（HS组）中观察到血管再生。在移植了2000胰岛当量、分别用HS或HSA培养且血糖水平和血糖正常动物百分比随时间相似的链脲佐菌素（STZ）诱导糖尿病裸鼠（n = 33）中测定胰岛功能。总之，在补充有HS的培养基中培养的人胰岛在体外显示出更高的存活率以及胰岛活力和功能。更高的体外存活率增加了可用于移植的胰岛数量。然而，当移植相似数量的HSA培养胰岛和HS培养胰岛时，对活力和功能的有益作用并未转化为改善的代谢演变。
