Highly Selective Intraportal Transplantation of Pancreatic Islets
Maciej JuszczakPaul KoonerKrystian PawelecGareth JonesStephen J. HughesAnila KumarStephen H. PowisMartin Press
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Pancreatic Islets
Islet cell transplantation
Objective To observe the effect of treating diabetic mice with microcapsulated rat islet cell transplantation.Methods Diabetic mice were randomly divided into 3 groups:control group,no-microencapsulated islet cells transplantation group and microencapsulated islet cells transplantation group.Normal saline,pared rat islet cells and microcapsulated islet cells were respectively transplantated into abdominal cavity of three groups of diabetic mice.Results The isolated islet cells had a good reaction for glucose stimulation.Both the microcapsulated islet cell transplantation and non-microcapsulated islet cell transplantation could be decreased the high blood glucose level,and the former one kept longer.Conclusion It is believed that microcapsulated islet cell transplantation exerts good effect on diabetic mice and the microcapsules have good immuno-isolating function.
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Objective To optimize the quantity of islet cells for homogeneous transplantation in type Ⅰdiabetes rats. Methods Homogeneous islet grafts were isolated from SD rats.The pancreas was gained by in situ perfusion,and then was digested by collagenase Ⅴ and cleared by Ficoll400 for islet cells. Islet cells were confirmed by DTZ staining and then calculated.AO-PI fluorescent staining was adopted to detect the activity of islet cells. Insulin release test was used to evaluate the function of islet cells.Type-Ⅰdiabetes rat models were induced by intravenous injection of STZ(60 mg/kg).The model rats were randomly divided into four groups according to the concentration of islet grafts:group A(6 000 IEQ/kg),group B(9 000 IEQ/kg),group C(12 000 IEQ/kg),group D(15 000 IEQ/kg).The levels of blood glucose and insulin were monitored at different time points. Results In group A and group B,blood glucose did not drop to the normal level after transplantation.In group C and group D,blood glucose reduced to the normal level after transplantation for 22 h,25 h,respectively.After islet transplantation for 3 d,the levels of insulin stepped up with the increased quantity of islet cells. Conclusion The concentration of 12 000 IEQ/kg is the optimized concentration for homogeneous transplantation of islet in rats.
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Background: Over the past 20 years, significant advances have been made in human islet transplantaiton. However, cases of prolonged insulin independence after islet allotransplantation have rarely been reported and over time, a slight, gradual decrease in insulin secretion appears to occur, as suggested by the lower C-peptide. Although preliminary clinical success achieved over the past few years has been considerably higher with whole pancreatic transplant than with isolated islet grafts, both approaches remain experimental. Islet grafts might gain, over time, increasing credibility and might eventually provide an easier alternative in terms of grafting procedures and patient management, as compared with the more traumatizing whole-pancreas transplantation. Also, using islet, re-transplantation is possible. But it is not known whether re-transplantation of islet could be suitable for those patients who lost grafted islet function. The aim of the present study was to investigate the benefits of re-transplantation of islet in previously simultaneous islets-kidney transplant(SIK) patient who have lost graft function. Methods: The recipient was a 32 year old male. First islet transplantation was underwent at December 25, 1999. However, the grafted islets lost function after 70 days. So we performed re-transplantation of islets. The isolation of islet was conducted sterilely on a laminar flow hood and isolated by a modified Recordi method. The islet was injected slowly into the liver via a cannular placed in the potal vein for 20 minutes. Results: Transplanted islets were 90,000 IEq at first islet transplantation, 370,000 IEq at second islet transplantation. The insulin requirement was reduced from 75-85 to 35-40 U/day, the basal C-peptide level was 1.5ng/mL at 7 days posttransplant Unfortunately, the grafted islets lost function after 70 days. After second transplantation, the insulin requirement was reduced to 26 U/day. Conclusions: Despite the continuous need for exogenous insulin therapy, islet transplantation can prevent wide glucose fluctuations, thus resulting in normalization of glycemic control and improvement in HbA1c, and also, show that islets can be successfully and safely re-transplanted intraportally in patients who have lost previously grafted islet function(J Kor Diabetes Asso 457∼466, 2000).
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Islet transplantation is an ideal option for the treatment of brittle diabetes. In this protocol, isolated and purified islets were transplanted via the hepatic portal vein into a blood group compatible recipient (Rickels and Robertson, Endocr Rev 40:631–668, 2019). In contrast to pancreas transplantation, this technique is less invasive and has fewer posttransplantation complications. Although the clinical outcomes of islet transplantation have improved dramatically after the Edmonton protocol was proposed, there is still much room for improvement. Islet transplantation in mice is one of the most commonly used models in islet transplantation studies. By implanting islets under the renal capsule, the rejection, survival, blood sugar fluctuations and C-peptide secretion of islets can be observed in real time. Our model is of great scientific and clinical significance for studying immune rejection in islet transplantation, improving the transplantation effect, and further prolonging the survival of transplanted islets (Shapiro et al., Nat Rev Endocrinol 13:268–277, 2017).
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Beta cell replacement is an exciting field where new beta cell sources and alternative sites are widely explored. The liver has been the implantation site of choice in the clinic since the advent of islet transplantation. However, in most cases, repeated islet transplantation is needed to achieve normoglycemia in diabetic recipients. This study aimed to investigate whether there are differences in islet survival and engraftment between a first and a second transplantation, performed 1 week apart, to the liver. C57BL/6 mice were accordingly transplanted twice with an initial infusion of syngeneic islets expressing green fluorescent protein (GFP). The second islet transplant was performed 1 week later and consisted of islets isolated from non-GFP C57BL/6-mice. Animals were sacrificed either 1 day or 1 month after the second transplantation. A control group received a saline infusion instead of GFP-expressing islets, 1 week later obtained a standard non-GFP islet transplant, and was subsequently sacrificed 1 month later. Islet engraftment in the liver was assessed by immunohistochemistry and serum was analyzed for angiogenic factors induced by the first islet transplantation. Almost 70% of islets found in the liver following repeated islet transplantation originated from the second transplantation. The vascular density in the transplanted non-GFP-expressing islets did not differ depending on whether their transplantation was preceded by a primary islet transplantation or saline administration only nor did angiogenic factors in serum prior to the transplantation of non-GFP islets differ between animals that had received a previous islet transplantation or a saline infusion. We conclude that first islet transplantation creates, by unknown mechanisms, favorable conditions for the survival of a second transplant to the liver.
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