Isolation and Purification of Adult Tibet Minipig Pancreas Islets in vitro

doi:10.3969/j.issn.1674-5817.2013.02.009

Abstract

Abstract: Objectives Identify the optimized techniques for the isolation and purification of pancreas islets to supply high quality islets for xenotransplantation in the treatment of diabetes. Methods Compare the quantities and effects of different mass concentration (1 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml) and the same mass concentration conditions at different time points (0 min, 15 min, 30 min, 45 min, 60 min, 75 min, 90 min) on Tibet minipig pancreas islets isolation. Compare the quantities and effects of Ficoll400 density gradient centrifugation, lymphatic separation liquid centrifugal, centrifugal, and placing the 4 ways on purification of islets. With AO/PI dyeing calculation islet cell survival rate. Results The best time points of different mass concentration collagen enzyme to acquire islets were confirmed, 1 mg/ml and 2 mg/ml groups for 45 min,3 mg/ml and 4 mg/ml groups for 30 min respectively. Among the 4 best time points, 1 mg/ml group gets the most amount of islets . Each group gets islets of complete morphology at best time point respectively. Islets number by many to less is placing> Ficoll400 density gradient centrifugation>lymphatic separation liquid centrifugal>centrifugal among 4 kinds of purification ways. The AO/PI staining after testing the survival rate of pancreatic islet cells for more than 95.00%. Conclusions In the condition of cutting tissue into pieces, collagen enzyme mass concentration 1 mg/ml, 45 min of digesting time can get more islets, Ficoll400 density gradient centrifugation collects islets most effectively.

Key words: Tibet minipig, Pancreas islets, Isolation, Purification

CLC Number:

Q95-33

QI Wen-jun, WANG Yi-nan, HU Qiang, TANG Hua, NASHUN Ba-ya-er, GU Wei-wang. Isolation and Purification of Adult Tibet Minipig Pancreas Islets in vitro[J]. Laboratory Animal and Comparative Medicine, 2013, 33(2): 126-129.

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