Lee R H, Carter J, Szot G L, Posselt A, Stock P
Transplantation Research Laboratory, University of California-San Francisco, San Francisco, California, USA.
Transplant Proc. 2008 Mar;40(2):384-6. doi: 10.1016/j.transproceed.2008.02.016.
Human islet transplant protocols frequently include a brief period of islet culture before transplantation. Some investigators have suggested that medium supplementation with human serum might quench collagenase activity and provide better culture conditions when compared with human albumin. We studied the effect of whole serum on islet count, islet equivalence, insulin secretion, and DNA content in human islets.
Adult human islets isolated from a single pancreas with purity >50% were cultured in identical 150 islet equivalent samples at 37 degrees C using CMRL 1066-based islet medium (Mediatech) supplemented with either 0.5% human albumin or 10% human AB serum. Prior to culture and after 3 days, islets were assessed in vitro using dithizone staining (n = 4), insulin release after static glucose stimulation (n = 8), and DNA content (n = 8).
After 3 days, islet mass (defined by the number of islets and islet equivalents counted after dithizone staining) was better preserved in islets cultured in 0.5% human albumin. Although the stimulation index and total DNA content were similar between groups, islets cultured in human albumin demonstrated greater absolute insulin secretion (p = .02) and insulin secretion per cell (p = .02).
When used to supplement CMRL 1066-based islet culture medium, human albumin preserves islet mass and secretory capacity better than whole human serum. Human serum offers no advantage in islet preservation or function.
人类胰岛移植方案通常包括移植前短暂的胰岛培养期。一些研究人员认为,与人类白蛋白相比,在培养基中添加人血清可能会抑制胶原酶活性并提供更好的培养条件。我们研究了全血清对人胰岛的胰岛数量、胰岛当量、胰岛素分泌和DNA含量的影响。
从单个胰腺分离出纯度>50%的成人人类胰岛,在37℃下使用基于CMRL 1066的胰岛培养基(Mediatech),以相同的150个胰岛当量样本进行培养,该培养基分别添加0.5%的人类白蛋白或10%的人类AB血清。在培养前和培养3天后,使用双硫腙染色(n = 4)、静态葡萄糖刺激后的胰岛素释放(n = 8)和DNA含量(n = 8)对胰岛进行体外评估。
3天后,在添加0.5%人类白蛋白的培养基中培养的胰岛,其胰岛质量(通过双硫腙染色后计数的胰岛数量和胰岛当量定义)得到了更好的保存。尽管两组之间的刺激指数和总DNA含量相似,但在人类白蛋白中培养的胰岛显示出更高的绝对胰岛素分泌量(p = 0.02)和每细胞胰岛素分泌量(p = 0.02)。
当用于补充基于CMRL 1066的胰岛培养基时,人类白蛋白在保存胰岛质量和分泌能力方面比全人血清更好。人血清在胰岛保存或功能方面没有优势。