Datar S P, Bhonde R R
S P College, Tilak Road, Pune-411030, India.
Cryo Letters. 2010 Nov-Dec;31(6):485-92.
The avian endocrine pancreas shares some similarities with those of mammals. Previously we have reported a technique of isolation of B islets from chick pancreas and also demonstrated their possible use for hypoglycemic drug screening as efficiently as mammalian islets. Here we describe a novel cryopreservative medium for the cryopreservation of chick islets. Isolated chick islets were suspended in a cryo medium consisting of Dulbecco's modified Minimum Essential Medium: Ham's F12 (1:1), Fetal Bovine Serum (20%), Dimethylsulfoxide (10%) with different concentrations (50 microg/ml to 500 microg/ml) of riboflavin or nicotinamide. The viability of revived islets after three and half months was checked by trypan blue dye exclusion and functionality was assessed by insulin secretion in response to glucose challenge. The maximum recovery of viable islets and insulin secretion was obtained in response to glucose challenge at 250 microg/ml concentration of Riboflavin or Nicotinamide. This is a first report on cryopreservation of chick islets exhibiting cryoprotective role of water soluble vitamins without vitamin C.
禽类的内分泌胰腺与哺乳动物的内分泌胰腺有一些相似之处。此前我们报道了一种从鸡胰腺中分离B胰岛的技术,并且还证明了它们在低血糖药物筛选中的应用潜力,其效率与哺乳动物胰岛相当。在此,我们描述了一种用于鸡胰岛冷冻保存的新型冷冻保存培养基。将分离出的鸡胰岛悬浮于一种冷冻培养基中,该培养基由杜氏改良Eagle培养基:哈姆F12(1:1)、胎牛血清(20%)、二甲基亚砜(10%)以及不同浓度(50微克/毫升至500微克/毫升)的核黄素或烟酰胺组成。通过台盼蓝拒染法检查复苏三个月半后的胰岛活力,并通过对葡萄糖刺激的胰岛素分泌来评估其功能。在核黄素或烟酰胺浓度为250微克/毫升时,对葡萄糖刺激的反应中,活胰岛和胰岛素分泌的回收率最高。这是关于鸡胰岛冷冻保存的首次报道,展示了不含维生素C的水溶性维生素的冷冻保护作用。