Down-regulation of proliferation does not affect the secretory function of transformed β-cell lines regardless of their anatomical configuration

Reers, Christina; Hauge-Evans, Astrid C.; Morgan, Noel G.; Persaud, Shanta J.; Jones, Peter M.

doi:10.4161/isl.3.3.15428

Cited by 9 publications

(5 citation statements)

References 30 publications

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“…β cells require homotypic cell-cell interaction to function correctly in vitro. Therefore, most studies use pure populations of insulin-secreting cells to generate functional pseudoislets [ 13 , 38 ]. It was shown that homotypic interactions within MIN6 PIs in which cells are assembled in a three-dimensional configuration were sufficient to significantly enhance secretory responsiveness to nutrients and non-nutrient stimuli when compared to equivalent cells configured as monolayers [ 12 ].…”

Section: Resultsmentioning

confidence: 99%

Formation of βTC3 and MIN6 Pseudoislets Changes the Expression Pattern of Gpr40, Gpr55, and Gpr119 Receptors and Improves Lysophosphatidylcholines-Potentiated Glucose-Stimulated Insulin Secretion

Drzazga

Cichońska

Koziołkiewicz

et al. 2020

Cells

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The impaired spatial arrangement and connections between cells creating islets of Langerhans as well as altered expression of G protein-coupled receptors (GPCRs) often lead to dysfunction of insulin-secreting pancreatic β cells and can significantly contribute to the development of diabetes. Differences in glucose-stimulated insulin secretion (GSIS) are noticeable not only in diabetic individuals but also in model pancreatic β cells, e.g., βTC3 and MIN6 β cell lines with impaired and normal insulin secretion, respectively. Now, we compare the ability of GPCR agonists (lysophosphatidylcholines bearing fatty acid chains of different lengths) to potentiate GSIS in βTC3 and MIN6 β cell models, cultured as adherent monolayers and in a form of pseudoislets (PIs) with pancreatic MS1 endothelial cells. Our aim was also to investigate differences in expression of the GPCRs responsive to LPCs in these experimental systems. Aggregation of β cells into islet-like structures greatly enhanced the expression of Gpr40, Gpr55, and Gpr119 receptors. In contrast, the co-culture of βTC3 cells with endothelial cells converted the GPCR expression pattern closer to the pattern observed in MIN6 cells. Additionally, the efficiencies of various LPC species in βTC3-MS1 PIs also shifted toward the MIN6 cell model.

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Section: Resultsmentioning

confidence: 99%

Formation of βTC3 and MIN6 Pseudoislets Changes the Expression Pattern of Gpr40, Gpr55, and Gpr119 Receptors and Improves Lysophosphatidylcholines-Potentiated Glucose-Stimulated Insulin Secretion

Drzazga

Cichońska

Koziołkiewicz

et al. 2020

Cells

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“…Luther et al showed an overall increase in β cell death which was manifested by reduced proliferation and increased apoptosis of MIN6 cells in PIs when compared to monolayers [23]. Reers et al showed that MIN6 PIs exhibit an overall reduction in β cell proliferation [31]. These changes in β cell viability and proliferation represent a limitation for the use of PIs in the study of β cell physiology.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Formation of β Cell Pseudoislets Using Islet-Derived Endothelial Cells

et al. 2013

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β cell pseudoislets (PIs) are used for the in vitro study of β-cells in a three-dimensional (3-D) configuration. Current methods of PI induction require unique culture conditions and extensive mechanical manipulations. Here we report a novel co-culture system consisting of high passage β-cells and islet-derived endothelial cells (iECs) that results in a rapid and spontaneous formation of free-floating PIs. PI structures were formed as early as 72 h following co-culture setup and were preserved for more than 14 d. These PIs, composed solely of β-cells, were similar in size to that of native islets and showed an increased percentage of proinsulin-positive cells, increased insulin gene expression in response to glucose stimulation, and restored glucose-stimulated insulin secretion when compared to β-cells cultured as monolayers. Key extracellular matrix proteins that were absent in β-cells cultured alone were deposited by iECs on PIs and were found in and around the PIs. iEC-induced PIs are a readily available tool for examining β cell function in a native 3-D configuration and can be used for examining β-cell/iEC interactions in vitro.

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“…The AUC for both pseudoislets groups was markedly smaller than that of the TCPS group, and similar to that of the normal mice group. Regarding the possibility of cell proliferation of cell clusters attribute to the improvement of IPGTT, both Kelly et al 12 and Reers et al 41 reported that the cell proliferation of pseudoislets was decreased compared with monolayers, which eliminate this concern. Besides, relative to TCPS group, mice in both pseudoislets groups also had higher serum insulin level.…”

Section: Discussionmentioning

confidence: 99%

Investigating the suspension culture on aggregation and function of mouse pancreatic β‐cells

Yang

et al. 2013

J Biomedical Materials Res

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The integrity and hierarchical structure of islet influence β-cells physiology dramatically. A culture substrate which can maintain or improve β-cells aggregation shall benefit cell therapy for diabetics. In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine β-cell line, MIN-6. The formation and biological performances of pseudoislets were investigated. Results showed that β-cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100-150 μm with high survival rate, while large pseudoislets (>250 μm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose-stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin-producing cells can be cultured on type IV collagen or Lipidure dishes to improve/maintain integrity prior to transplantation.

show abstract

Down-regulation of proliferation does not affect the secretory function of transformed β-cell lines regardless of their anatomical configuration

Cited by 9 publications

References 30 publications

Formation of βTC3 and MIN6 Pseudoislets Changes the Expression Pattern of Gpr40, Gpr55, and Gpr119 Receptors and Improves Lysophosphatidylcholines-Potentiated Glucose-Stimulated Insulin Secretion

Formation of βTC3 and MIN6 Pseudoislets Changes the Expression Pattern of Gpr40, Gpr55, and Gpr119 Receptors and Improves Lysophosphatidylcholines-Potentiated Glucose-Stimulated Insulin Secretion

In Vitro Formation of β Cell Pseudoislets Using Islet-Derived Endothelial Cells

Investigating the suspension culture on aggregation and function of mouse pancreatic β‐cells

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