摘要
目的建立新型成人胰岛细胞分离纯化方法,观察成人胰岛细胞移植的安全性与有效性。方法对14例1型糖尿病(T1DM)患者PFC与UW液双层冷藏胰腺,Liberase酶消化,COBE2991型专用胰岛细胞分离机分离及连续密度梯度纯化,获取高纯度与高活性的胰岛细胞。采用外科方法,将短期培养的胰岛细胞经门静脉移植到肝脏内。术后监测血糖与胰岛素用量、C肽与糖化血红蛋白水平以及肝肾功能,采用t检验与术前比较差异的统计学意义。结果 47个胰腺成功分离胰岛细胞,平均数量32.5万IEQ、纯度81.4﹪、活率93.2﹪、刺激指数3.26,病原学结果均阴性。14例T1DM患者实施成人胰岛细胞移植25次,每次移植平均胰岛数量为10200IEQ/kg。采用无激素免疫抑制治疗。随访18个月至5年,8例完全撤除胰岛素,6例胰岛素用量较术前减少60﹪以上。术后血糖稳定维持在正常水平,C肽均超过0.5ng/ml,糖化血红蛋白基本正常,肝肾功能稳定维持正常,无胰岛移植并发症。结论新型成人胰岛细胞分离纯化方法可靠,成人胰岛细胞移植治疗1型糖尿病临床疗效较好,手术简捷,安全性好。
Objective To establish a new technique for isolating pancreatic islets of langerhans and evaluate its clinical efficacy and safety. Methods Pancreases were stored using the ＂2-layer method＂ of the oxygenated perfluorochemical （PFC） and UW solution. The pancreases were digested by Liberase collagenase enzyme and islets were purified using continuous gradients of Ficoll-diatrizoic acid on a refrigerated COBE 2991 centrifuge. Cultured islets were infused by surgical approach to the liver via portal vasculature. Levels of blood glucose, C-peptide and HbAlc were evaluated. Dose of insulin, liver and renal functions after transplantation were compared with those before transplantation. Results Islets of langerhans were isolated successfully in 47 pancreases. The average islet yield was 325 000 islet equivalents （IEQ）. Islet purity and viability were 81.4 % and 93.2 %, respectively. The stimulation index （SI） of human islet was 3.26. No viruses or bacteria were detected in each islet preparation. Twenty-five islet infusions have been carried out in 14 subjects with typeldiabetes mellitus （T1DM）. The average islet mass for infusion was 10 200 IEQ/kg. The glucocorticoid-free immunosuppressive regimen was used. During the follow-up period （18 months to 5 years）, 8 recipients remained insulin-independent. The dosage of insulin decreased by 60 % in 6 patients. The levels of blood glucose and HbAlc were all within normal range and liver and renal functions were normal. C-peptide level was above 0.5 ng/ml. No complications related to islet infusion was observed. Conclusions The new technique is more efficient for isolating pancreatic islet of langerhans. Islet transplantation is effective and safe for treatingT1DM.
出处
《中华细胞与干细胞杂志（电子版）》
2011年第2期37-41,共5页
Chinese Journal of Cell and Stem Cell（Electronic Edition）
基金
国家自然科学基金(30571759)
全军"十五"重大课题(04Z007)
全军"十一五"计划课题(06G037)
福建省科技厅重大攻关课题(2002Y007)
关键词
糖尿病
成人胰岛细胞
移植
Diabetes Mellitus
Adult islet
Transplantation
