Intrapatient Variations in Type 1 Diabetes-specific iPS Cell Differentiation Into Insulin-producing Cells

Thatava, Tayaramma; Kudva, Yogish C.; Edukulla, Ramakrishna; Squillace, Karen A.; Lamo, Josep Genebriera De; Khan, Yulia; Sakuma, Toshie; Ohmine, Seiga; Terzic, André; Ikeda, Yasuhiro

doi:10.1038/mt.2012.245

Cited by 92 publications

(76 citation statements)

References 53 publications

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“…Thatava and coworkers generated iPS cells from a diabetic patient and differentiated these cells to insulin producing cells. 26 We also produced insulin producing cells from human IPS cells after 23 days with bFGF and nicotinamide. 20 .…”

Section: Discussionmentioning

confidence: 99%

Untitled

Shaer

Azarpira

Karimi³

et al. 2016

Exp Clin Transplant

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Objectives: Diabetes results from inadequate insulin production from pancreatic ß-cells. Islet cell replacement is an effective approach for diabetes treatment; however, it is not sufficient for all diabetic patients. Thus, finding a new source with effective maturation of ß-cells is the major goal of many studies. MicroRNAs are a class of small noncoding ribonucleic acid that regulate gene expression through posttranscriptional mechanisms. MicroRNA-7 has high expression level during pancreatic islet development in humans, thereby playing a critical role in pancreatic β-cell function. We study aimed to develop a protocol to differentiate human-induced pluripotent stem cells efficiently into isletlike cell clusters in vitro by using microRNA-7. Materials and Methods: Human-induced pluripotent stem cell colonies were transfected with hsamicroRNA-7 by using siPORT NeoFX transfection agent. Total ribonucleic acid was extracted 24 and 48 hours after transfection. The expression of transcription factors which were important during pancreases development was also performed. On the third day, the potency of the clusters was assessed in response to high glucose levels. Diphenylthiocarbazone was used to identify the existence of the ß-cells. The presence of insulin and Neurogenin-3 proteins was investigated by immunocytochemistry. Results: Morphologic changes were observed on the first day after chemical transfection, and cell clusters were formed on the third day. The expression of pancreatic specific transcription factors was increased on the first day and significantly increased on the second day. The isletlike cell clusters were positive for insulin and Neurogenin-3 proteins in immunocytochemistry. The clusters were stained with Diphenylthiocarbazone and secreted insulin in a glucose challenge test. Conclusions: MicroRNA-7 transcription factor net-work is important in pancreatic endocrine differe-ntiation. Chemical transfection with microRNA-7 can differentiate human induced pluripotent stem cells into functional isletlike cell clusters in a short time.

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Section: Discussionmentioning

confidence: 99%

Untitled

Shaer

Azarpira

Karimi³

et al. 2016

Exp Clin Transplant

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“…The authors believe that this method allows reproducible generation of genomic modification-free induced pluripotent stem cells from diabetic patients for autologous cell replacement therapy. 47 The virusmediated delivery of reprogramming factors, such as c-Myc and KLF4, results in permanent integration of these oncogenes with subsequent permanent genetic alterations. [48][49] Therefore, various methods have been used to generate induced pluripotent stem cells with lower genetic integration such as mRNAs, plasmid transient transfection, episomal vector, nonintegrating Sendi virus, and adenovirus.…”

Section: Discussionmentioning

confidence: 99%

Untitled

Shaer

Azarpira

Vahdati³

et al. 2015

Exp Clin Transplant

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Objectives: In diabetes mellitus type 1, beta cells are mostly destroyed; while in diabetes mellitus type 2, beta cells are reduced by 40% to 60%. We hope that soon, stem cells can be used in diabetes therapy via pancreatic beta cell replacement. Induced pluripotent stem cells are a kind of stem cell taken from an adult somatic cell by "stimulating" certain genes. These induced pluripotent stem cells may be a promising source of cell therapy. This study sought to produce isletlike clusters of insulin-producing cells taken from induced pluripotent stem cells. Materials and Methods: A human-induced pluripotent stem cell line was induced into isletlike clusters via a 4-step protocol, by adding insulin, transferrin, and selenium (ITS), N2, B27, fibroblast growth factor, and nicotinamide. During differentiation, expression of pancreatic β-cell genes was evaluated by reverse transcriptase-polymerase chain reaction; the morphologic changes of induced pluripotent stem cells toward isletlike clusters were observed by a light microscope. Dithizone staining was used to stain these isletlike clusters. Insulin produced by these clusters was evaluated by radio immunosorbent assay, and the secretion capacity was analyzed with a glucose challenge test. Results: Differentiation was evaluated by analyzing the morphology, dithizone staining, real-time quantitative polymerase chain reaction, and immunocytochemistry. Gene expression of insulin, glucagon, PDX1, NGN3, PAX4, PAX6, NKX6.1, KIR6.2, and GLUT2 were documented by analyzing real-time quantitative polymerase chain reaction. Dithizonestained cellular clusters were observed after 23 days. The isletlike clusters significantly produced insulin. The isletlike clusters could increase insulin secretion after a glucose challenge test. Conclusions:This work provides a model for studying the differentiation of human-induced pluripotent stem cells to insulin-producing cells.

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“…After the islets are seeded in the scaffold, it will then be attached to the omentum using thrombin surrounding which an omental 'pouch' will be created to secure the implant. However, it is worth noting that this 'scaffold' is not immunoprotective, and the patient will require conventional immunosuppression to prevent graft rejection [184].…”

Section: Biohub -Drimentioning

confidence: 99%