Chadwick D R, Robertson G S, Rose S, Contractor H, James R F, Bell P R, London N J
Department of Surgery, Leicester Royal Infirmary, United Kingdom.
Transplantation. 1993 Aug;56(2):288-93. doi: 10.1097/00007890-199308000-00006.
The aim of this study was to establish the beneficial effect of storage of pancreatic digest in University of Wisconsin solution on porcine islet purification, the mechanism of this effect, and the components of UW responsible. Ten porcine pancreata were collagenase-digested, and samples of digest were washed and stored for 1 hr in either UW or minimum essential medium at 4 degrees C, prior to separation on continuous linear density gradients of bovine serum albumin. Samples of digest from a further ten pancreata were similarly treated, comparing storage in MEM, UW, and five solutions varying in lactobionate:chloride ratio and raffinose content. The purity of the islet preparations and the densities of islets and exocrine tissue were determined from insulin and amylase assay of aliquots aspirated from these gradients. Washing and storage of digest in UW markedly improved islet purity, compared with MEM, due to an increase in the density of exocrine tissue. Exocrine tissue density following storage was dependent upon the control of acinar cell volume, rather than exocrine enzyme discharge, and was determined primarily by the chloride:lactobionate ratio of the storage solution. Raffinose was of little additional benefit, while the beneficial effect of UW was greater than that due to its lactobionate and raffinose content alone. In conclusion, inadequate purification of islets results from exocrine tissue swelling. This swelling is reduced by storage of the pancreatic digest in UW solution, due primarily to the replacement of chloride by lactobionate in UW.
本研究的目的是确定将胰腺消化物储存在威斯康星大学溶液中对猪胰岛纯化的有益作用、这种作用的机制以及威斯康星大学溶液中起作用的成分。将10个猪胰腺用胶原酶消化，消化物样品在4℃下用威斯康星大学溶液或最低限度基本培养基洗涤并储存1小时，然后在牛血清白蛋白连续线性密度梯度上进行分离。对另外10个胰腺的消化物样品进行类似处理，比较在最低限度基本培养基、威斯康星大学溶液以及5种乳糖酸盐：氯化物比例和棉子糖含量不同的溶液中的储存情况。从这些梯度中吸出的等分试样通过胰岛素和淀粉酶测定来确定胰岛制剂的纯度以及胰岛和外分泌组织的密度。与最低限度基本培养基相比，用威斯康星大学溶液洗涤和储存消化物可显著提高胰岛纯度，这是由于外分泌组织密度增加所致。储存后外分泌组织密度取决于腺泡细胞体积的控制，而非外分泌酶的释放，并且主要由储存溶液的氯化物：乳糖酸盐比例决定。棉子糖几乎没有额外益处，而威斯康星大学溶液的有益作用大于仅因其乳糖酸盐和棉子糖含量所产生的作用。总之，胰岛纯化不充分是由于外分泌组织肿胀所致。将胰腺消化物储存在威斯康星大学溶液中可减轻这种肿胀，这主要是因为威斯康星大学溶液中的氯化物被乳糖酸盐取代。
