title: Facile mechanical shaking method is an improved isolation approach for islet preparation and transplantation.
abstract: Successful islet isolation is crucial for islet transplantation and cell treatment for type 1 diabetes. Current isolation methods are able to obtain 500-1,000 islets per rat, which results in a waste of ≥50% of total islets. In the present study, a facile mechanical shaking method for improving islet yield (up to 1,500 per rat) was developed and summarized, which was demonstrated to be more effective than the existing well-established stationary method. The present results showed that isolated islets have a maximum yield of 1,326±152 when shaking for 15 min for the fully-cannulated pancreas. For both fully-cannulated and half-cannulated pancreas in the presence of rat DNAse inhibitor, the optimal shaking time was amended to 20 min with a further increased yield of 1,344±134 and 1,286±124 islets, respectively. Furthermore, the majority of the isolated islets were morphologically intact with a well-defined surface and almost no central necrotic zone, which suggested that the condition of islets obtained via the mechanical shaking method was consistent with the stationary method. Islet size distribution was also calculated and it was demonstrated that islets from the stationary method exhibited the same size distribution as the non-cannulated group, which had more larger islets than the fully-cannulated and half-cannulated groups isolated via the shaking method. In addition, the results of glucose challenge showed that the refraction index of each group was >2.5, which indicated the well-preserved function of isolated islets. Furthermore, the transplanted islets exhibited a therapeutic effect after 1 day of transplantation; however, they failed to control blood glucose levels after ~7 days of transplantation. In conclusion, these results demonstrated that the facile mechanical shaking method may markedly improve the yield of rat islet isolation, andin vitroandin vivoinvestigation demonstrated the well-preserved function of isolated islets in the control of blood glucose. Therefore, the facile mechanical shaking method may be an alternative improved procedure to obtain higher islet yield for islet preparation and transplantation in the treatment of type 1 diabetes.
authors.affiliation: Department of Anatomy, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Nina
authors.initials: N
authors.lastname: Yin
authors.affiliation: Department of Anatomy, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.affiliation: Department of Anatomy, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Anatomy, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Anatomy, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Fei
authors.initials: F
authors.lastname: Yan
authors.affiliation: Department of Anatomy, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Zheng
authors.affiliation: Department of Acupuncture and Moxibustion, College of Acupuncture and Orthopedics, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China; Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
authors.forename: Zebin
authors.initials: Z
authors.lastname: Chen
keywords: islet isolation; islet transplantation; mechanical shaking method; type 1 diabetes