Effects of Cryopreservation on in Vitro and in Vivo Long-Term Function of Human Islets1

Piemonti, Lorenzo; Bertuzzi, Federico; Nano, Rita; Leone, Biagio Eugenio; Socci, C.; Pozza, G.

doi:10.1097/00007890-199909150-00011

Cited by 29 publications

(15 citation statements)

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“…The reduced insulin release on day 21 may be attributed to inappropriate culture conditions. Piemonti et al reported the loss of b cell function in cryopreserved islets Improvement of Human Islet Cryopreservation by a p38 MAPK Inhibitor within three months (44). We did not observe the decline of b cell function following transplantation.…”

Section: Omori Et Alcontrasting

confidence: 69%

Improvement of Human Islet Cryopreservation by a p38 MAPK Inhibitor

Omori

Valiente

Orr

et al. 2007

American Journal of Transplantation

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The activation of p38 mitogen-activated protein kinase (MAPK) has been shown to cause ischemia/reperfusion injury of several organs used for transplantation and also to play a significant role in primary islet graft nonfunction. Activation of p38 MAPK may also occur during islet cryopreservation and thawing. In this study, a p38 MAPK inhibitor (p38IH) was applied to human islet cryopreservation to improve islet yield and quality after thawing. Under serum-free conditions, human islets were cryopreserved, thawed and cultured using our standard procedures. Three types of solutions were tested: conventional RPMI1640 medium (RPMI), a newly developed islet cryopreservation solution (ICS), and ICS supplemented with a p38IH, SD-282 (ICS-p38IH). Activation or inhibition of p38 MAPK was demonstrated by the diminished phosphorylation of HSP27 substrate. Islet recovery on day 2 after thawing was highest with ICS-p38IH and islet viability was not significantly different in the three groups. b Cell numbers and function were the highest in islets cryopreserved with ICS-p38IH. Glucosestimulated human C-peptide levels were 86% of that of the nonfrozen islets when measured 4 weeks after transplantation into NODscid mice. This improvement may provide an opportunity to establish islet banks and allow the use of cryopreserved islets for clinical transplantation.

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Section: Omori Et Alcontrasting

confidence: 69%

Improvement of Human Islet Cryopreservation by a p38 MAPK Inhibitor

Omori

Valiente

Orr

et al. 2007

American Journal of Transplantation

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“…Interestingly, human and porcine islets with higher levels of insulin secretion at both the basal level and stimulated level of glucose stimulation were less likely to reverse diabetes in nude mouse transplantations, whereas a 100% cure rate was achieved by NHP islets. If islets release high amount of insulin at basal stimulus indicates unhealthy status of islets (6-8). …”

Section: Resultsmentioning

confidence: 99%

Factors Affecting Transplant Outcomes in Diabetic Nude Mice Receiving Human, Porcine, and Nonhuman Primate Islets

Loganathan¹,

Graham²,

Radosevich³

et al. 2013

Transplantation

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Background In the absence of a reliable islet potency assay, nude mice transplant is the criterion standard to assess islet quality for clinical transplantation. There are factors other than islet quality that affect the transplant outcome. Methods Here, we analyzed the transplant outcomes in 335 nude mice (NM) receiving islets from human (n=103), porcine (n=205), and non-human primate (NHP) donors (n=27). The islets (750, 1000, and 2000 islet equivalents) were transplanted under the kidney capsule of streptozotocin (STZ) induced diabetic NM. Results The proportion of mice that achieved normoglycemia was significantly higher in the group implanted with 2000 IEQ of human, porcine, or NHP islets (75% normoglycemic) versus groups that were implanted with 750 IEQ (7% normoglycemic) and 1000 IEQ (30% normoglycemic). In this study, we observed that the purity of porcine islet preparations (P ≤ .001), islet pellet size in porcine preparations (P ≤ .01) and mice recipient body weight for human islets preparations (P =.013), was independently associated with successful transplant outcome. NHP islets of 1000 IEQ were sufficient to achieve normoglycemic condition (83%). An islet mass of 2000 IEQ, high islet purity, increased recipient body weight, and high islet pellet volume increased the likelihood of successful reversal of diabetes in transplanted mice. Also, higher insulin secretory status of islets at basal stimulus was associated with a reduced mouse cure rate. The cumulative incidence of graft failure was significantly greater in human islets (56.12%) compared with porcine islets 35.57% (P ≤ .001). Conclusion Factors affecting NM bioassay were identified (islet mass, islet purity, pellet size, in vitro insulin secretory capability and mouse recipient body weight) and should be considered when evaluating islet function.

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“…The islets (250 islet/ml) were cultured freeEvaluation of Graft Function floating (37°C, 5% CO 2 ) in RPMI-1640 medium (BioWhittaker, Walkersville, MD, USA) supplemented with Blood sugar levels were measured 15, 30, and 60 min L-glutamine (Sigma), penicillin and streptomycin (1000 after the end of surgical procedure, daily for the first U/ml and 10 mg/ml; Sigma), and 10% (v/v) fetal calf week and then every second day posttransplantation. serum (HyClone, Celbio, Logan, UT, USA) for [20][21][22][23][24] Surgical death was defined as death within the first 7 h before the transplant. Islet purity was >90%.…”

Section: Methodsmentioning

confidence: 99%

Intrahepatic Islet Transplant in the Mouse: Functional and Morphological Characterization

Melzi¹,

Sanvito²,

Mercalli³

et al. 2008

Cell Transplant

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Although in a clinical setting islet transplantation is normally performed by percutaneous intrahepatic infusion, the kidney capsule has been the site of choice in nearly all the studies using mice. In the present study, we extensively characterized the mouse model of intraportally transplanted islets with the purpose to propose it as a model to study islet transplantation. C57BL/6 (n = 78) and BALB/C (n = 53) recipients were transplanted with 400 autologous islets alternatively through the portal vein (PV-Tx) or under the kidney capsule (KC-Tx). Glucose concentration during the first hour after syngeneic islet infusion was associated with subsequent long-term function confirming that early events have long-term effects on graft function. In both strains tested the probability to achieve islet function was significantly lower for PV-Tx than KC-Tx. Also in allogeneic models (C57BL/6 to BALB/C, n = 104; BALB/C to C57BL/6, n = 77) the probability to achieve primary function was significantly lower for PV-Tx than KC-Tx and the site of transplantation significantly affected the graft survival. Histological evaluation of livers showed the presence of features (embolism, thrombosis, focal areas of liver necrosis) that are absent in the kidney subcapsular site. Finally, significant differences in the outcome of PV-Tx were observed between the Th type 1 inflammatory-prone C57BL/6 mouse and the type 2 inflammatory-prone BALB/C mouse. Intraportal islet graft model has some features that are more similar to human clinical islet transplantation and should be used as a model to study not only engraftment but also mechanisms of immune suppression and immune tolerance. INTRODUCTIONing engraftment in the liver (5) has been suggested to be a prime factor necessitating the very large number of islets needed to achieve normoglycemia (2). InvestigaRodent models have been widely used to study the causal mechanisms of failure of islet transplantation and tion of the mechanisms of islet cell loss during intahepatic islet transplantation and of ways to prevent their loss to test protocols aimed at preventing islet graft failure. In most rodent models, islets are transplanted under the would seem important (24,27). Yet, few studies in murine models address this be means of intrahepatic transkidney capsule, whereas islet transplantation in humans is performed by percutaneous intrahepatic infusion plantation. A systematic study of the intrahepatic islet transplant in the mouse model is also lacking. through the portal vein (25). Intrahepatic islet infusion in humans is associated with an immediate blood-mediHere we describe allogenic and syngeneic intrahepatic islet transplantation models in two mouse strains, ated inflammatory reaction, thrombosis, and hepatic tissue ischemia (1,2,(4)(5)(6)13,18,22,23,28). Kidney subcapdescribe outcome in detail, and compare it with that of renal subcapsular islet transplantation. The findings sular transplantation is not intravascular and is devoid of these complications. Thus, although transplantation ...

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Effects of Cryopreservation on in Vitro and in Vivo Long-Term Function of Human Islets1

Abstract: These findings showed that cryopreservation affects the function of isolated human islets, maintaining in vivo function for a limited period of time.

Cited by 29 publications

References 50 publications

Improvement of Human Islet Cryopreservation by a p38 MAPK Inhibitor

Improvement of Human Islet Cryopreservation by a p38 MAPK Inhibitor

Factors Affecting Transplant Outcomes in Diabetic Nude Mice Receiving Human, Porcine, and Nonhuman Primate Islets

Intrahepatic Islet Transplant in the Mouse: Functional and Morphological Characterization

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