Isolation of Mouse Pancreatic Islets of Langerhans

Ramírez-Domínguez, Miriam

doi:10.1007/978-3-319-39824-2_3

Cited by 8 publications

(6 citation statements)

References 60 publications

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“…Although this method was very useful in medicine and was accompanied by success, there were major drawbacks to these methods due to some limitations (Cefalu & Kirkman, ). Later, isolation and transplantation of islets technique were developed to cure diabetes (Fotino, Vergani, Fiorina, & Pileggi, ; Ramirez‐Dominguez, ). However, the patients who received the islets survived no more than 5 years.…”

Section: Discussionmentioning

confidence: 99%

Therapeutic potential of human mesenchymal stem cells derived beta cell precursors on a nanofibrous scaffold: An approach to treat diabetes mellitus

Hoveizi

Tavakol

2018

Journal Cellular Physiology

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Diabetes mellitus is an autoimmune and chronic disorder that is rapidly expanding worldwide due to increasing obesity. In the current study, we were able to design a reliable 3‐dimensional differentiation process of human Wharton's jelly mesenchymal stem cells into pancreatic beta cell precursors (PBCPs) and detected that transplanted PBCPs could improve hyperglycemia in a diabetes‐induced model in mice. Polylactic acid/chitosan nanofibrous scaffold was prepared using an electrospinning method. Quantitative real‐time reverse transcription‐polymerase chain reaction and immunocytochemistry analysis were carried out to assess pancreatic marker expression in the differentiated cells. PBCPs were transplanted under the kidney capsule of diabetic mice that induced streptozotocin injection 14 days before the transplantation. Moreover, an intraperitoneal glucose tolerance test (ipGTT) was carried out 2 and 4 weeks after the transplantation to measure the reaction to a sudden increase of the blood glucose level in the transplanted animals. The results indicated that the expression of SRY (sex determining region Y)‐box (Sox17), forkhead box A2 (FoxA2), pancreatic and duodenal homeobox 1 (Pdx1), neurogenin 3 (Ngn3), hepatic nuclear factor 4, alpha (Hnf4α), and NK2 homeobox 2 (Nkx2.2) were increased significantly in the differentiated cells compared with that of the control group. In the current study, the diabetic disease was confirmed by measuring blood glucose and proved by conducting some other behavioral tests. After the PBCPs transplantation in a diabetic model, the ipGTT and hyperglycemia investigation during the determinant times confirmed the disease's significant improvement in the experimental models. In this study, some preclinical data suggested that the transplantation of PBCPs associated with appropriate nanofiber scaffold can be utilized for the treatment of diabetes models. In addition, studies are required to elucidate the molecular mechanism of PBCPs acting in diabetes models before being used for patients with diabetes.

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Section: Discussionmentioning

confidence: 99%

Therapeutic potential of human mesenchymal stem cells derived beta cell precursors on a nanofibrous scaffold: An approach to treat diabetes mellitus

Hoveizi

Tavakol

2018

Journal Cellular Physiology

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“…Pancreatic islets were isolated from mice with digestive enzyme ( 13 , 14 ). Briefly, a mouse was euthanized with CO 2 , followed by cervical dislocation, and then placed supine under a stereomicroscope, with the abdomen cleaned with 75% ethanol.…”

Section: Methodsmentioning

confidence: 99%

Hyperuricemia Predisposes to the Onset of Diabetes via Promoting Pancreatic β-Cell Death in Uricase-Deficient Male Mice

Lü¹,

He²,

Cui³

et al. 2020

Diabetes

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Clinical studies have shown a link between hyperuricemia (HU) and diabetes, while the exact effect of soluble serum urate on glucose metabolism remains elusive. This study aims to characterize the glucose metabolic phenotypes and investigate the underlying molecular mechanisms using a novel spontaneous HU mouse model in which the uricase ( Uox ) gene is absent. In an attempt to study the role of HU in glycometabolism, we implemented external stimulation on Uox knockout (KO) and wild-type (WT) males with a high-fat diet (HFD) and/or injections of multiple low-dose streptozotocin (MLD-STZ) to provoke the potential role of urate. Notably, while Uox- KO mice developed glucose intolerance in the basal condition, no mice spontaneously developed diabetes, even with aging. HFD-fed Uox- KO mice manifested similar insulin sensitivity compared with WT controls. HU augmented the existing glycometabolism abnormality induced by MLD-STZ and eventually led to diabetes, as evidenced by the increased random glucose. Reduced β-cell masses and increased terminal deoxynucleotidyl TUNEL-positive β-cells suggested that HU-mediated diabetes was cell death dependent. However, urate-lowering therapy (ULT) cannot ameliorate the diabetes incidence or reverse β-cell apoptosis with significance. ULT displayed a significant therapeutic effect of HU-crystal–associated kidney injury and tubulointerstitial damage in diabetes. Moreover, we present transcriptomic analysis of isolated islets, using Uox -KO versus WT mice and streptozotocin-induced diabetic WT (STZ-WT) versus diabetic Uox -KO (STZ-KO) mice. Shared differentially expressed genes of HU primacy revealed Stk17β is a possible target gene in HU-related β-cell death. Together, this study suggests that HU accelerates but does not cause diabetes by inhibiting islet β-cell survival.

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“…CMRL 1066 media was tested because it is clinically used for transplantation, and short-term cold preservation is practiced as it would prevent the shock to cells from the change from other preservation solution to transplant media 12 . Moreover, CMRL 1066 has been shown to prevent the growth of immune cells, such as dendritic and endothelial cells 37 . However, to investigate the difference between CMRL 1066 and UW preservation at 4°C, we performed a comparative analysis.…”

Section: Discussionmentioning

confidence: 99%

Assessment of Culture/Preservation Conditions of Human Islets for Transplantation

et al. 2022

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Islet culture before clinical transplantation has been adopted by various centers, but its effect on the survival and function of islets relative to the culture conditions and media needs further assessment. Human islets were cultured or preserved under four different conditions and three media options. Parameters such as recovery, viability, function, islet damage, and gene expressions for markers of hypoxia, and inflammation were assessed after 48-h culture or preservation. Preservation of islets was performed at 4°C in Connaught’s Medical Research Lab (CMRL) and University of Wisconsin (UW) media. Islets were cultured at 22°C, 37°C, and 37°C–22°C in CMRL and PRODO culture media. Islets preserved in UW solution had visually good morphology and exhibited higher recovery with less islet damage compared with the rest of the groups, whereas islets preserved in CMRL at 4°C resulted in poor morphology, recovery, viability, and function compared with the rest of the treatment conditions. Culture at 22°C and 37°C demonstrated an increase in the expression of inflammatory and hypoxia-related genes. In conclusion, islets preserved at 4°C in UW solution showed the best overall outcomes after 48 h compared with islets cultured at 22°C, 37°C, or 37°C–22°C in PRODO. Advancement in islet culture media is warranted to reduce inflammatory gene activation and improve recovery of islets for transplantation.

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Isolation of Mouse Pancreatic Islets of Langerhans

Cited by 8 publications

References 60 publications

Therapeutic potential of human mesenchymal stem cells derived beta cell precursors on a nanofibrous scaffold: An approach to treat diabetes mellitus

Therapeutic potential of human mesenchymal stem cells derived beta cell precursors on a nanofibrous scaffold: An approach to treat diabetes mellitus

Hyperuricemia Predisposes to the Onset of Diabetes via Promoting Pancreatic β-Cell Death in Uricase-Deficient Male Mice

Assessment of Culture/Preservation Conditions of Human Islets for Transplantation

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