摘要
目的:研究三维组织细胞旋转培养系统(RCCS)对胚胎胰岛冷冻保存复苏后质量的影响。方法:将胚胎胰岛平均分为实验组1、2和对照组,实验组1、2冷冻保存胰岛,冷冻保存前后分别用RCCS培养和普通培养,对照组经RCCS培养新鲜胰岛。切取胚胎胰腺,经胶原酶V消化、纯化,培养3d,然后进行标准冷冻保存,复苏后继续培养,并检测各组胰岛数量及活性。结果:纯化后每个胚胎收获胰岛最多5115.43IEQ,最少2331.98IEQ,平均(3551.27±253.76)IEQ。实验组1胰岛细胞存活率、胰岛素释放量、胰岛素刺激指数均高于实验组2。结论:RCCS培养有利于胰岛细胞的生长繁殖,使其具有更好的分泌胰岛素能力,该方法同胰岛冷冻保存相结合,可以进一步提高胰岛的冷冻保存效果,为胰岛库的成功建立探索出一条新的途径。
Objective：To study the impact of rotary cell culture system（RCCS） on the quality and activity of fetal islets of pancreas after cryopreservation and resuscitation.Methods：The fetal islets of pancreas were assigned into three groups averagely,experiment group 1,2 and control group.In experiment group1 and 2,fetal islets of pancreas were cultivated in RCCS and common culture respectively.While in control group,fresh fetal islets of pancreas were cultivated in RCCS all the time.We resected the fetal pancreas and digested with Collagenase V and purified with ficoll solution.Then standard cryopreservation step was adopted,and after resuscitation,the islets of pancreas were cultivated continuously.And the quantity,quality and activity of the islets of pancreas,and insulin stimulation test results in all groups were detected.Results：After purification,islets got from one fetal pancreas reached 2,331.98-5,115.43 IEQ（average 3,551.27± 253.76 IEQ） .The survival rate,the insulin release and stimulation index in islets of pancreas of group 1 incubated in RCCS were higher than those in common culture.Conclusion：RCCS are good for the growth and proliferation of islets of pancreas,and the islets of pancreas cultivated in RCCS have better insulin secreting capacity.RCCS combined standard cryopreservation method may further improve the cryopreservation effect.
出处
《中国现代普通外科进展》
CAS
2009年第10期846-849,共4页
Chinese Journal of Current Advances in General Surgery
关键词
失重
细胞培养技术
胰岛移植
低温保存
胚胎胰腺
Weightlessness·Cell culture techniques·Islets of langerhans transplantation·Cryopreservation·Fetal pancreas
