Transcriptomics and single-cell RNA sequencing analysis of long-term cultured human islets in vitro [RNA-Seq]
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ABSTRACT:
PROVIDER: PRJNA1181775 | ENA |
REPOSITORIES: ENA
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Project description:β-cell dysfunction in pancreatic islets is the leading cause of progression to type I diabetes (T1D), but the reason for the loss of β-cell mass in T1D is still not fully understood. Using in vitro long-term cultured (LTC) human islets, we have investigated gene profiling and subtype cells changes in the leftover cells and compared them with residual cells in samples of T1D or T2D. While most of the dysregulated genes were downregulated during LTC, we found there are specific groups of mRNA or miRNA that were upregulated, and they are involved in specific pathways. In general, α-cells and β-cells of LTC-islets have elevated expressions of MAFB and MAFA, respectively. We also found exocrine cells were eliminated faster than endocrine cells, and β-cells were lost at a higher rate than α-cells. Interestingly, a specific group of cells that were classified as α-cells but have immature β-cells-like characteristics, were enriched in LTC, revealed the possibility of transdifferentiating of α-cells to β-cells under in vitro culture. Our results suggested that there are intrinsic cellular and molecular mechanisms in pancreatic cells that are associated with their maturity and correlated with their survival ability under unfavorable living conditions.
2025-01-15 | GSE280983 | GEO
Project description:β-cell dysfunction in pancreatic islets is the leading cause of progression to type I diabetes (T1D), but the reason for the loss of β-cell mass in T1D is still not fully understood. Using in vitro long-term cultured (LTC) human islets, we have investigated gene profiling and subtype cells changes in the leftover cells and compared them with residual cells in samples of T1D or T2D. While most of the dysregulated genes were downregulated during LTC, we found there are specific groups of mRNA or miRNA that were upregulated, and they are involved in specific pathways. In general, α-cells and β-cells of LTC-islets have elevated expressions of MAFB and MAFA, respectively. We also found exocrine cells were eliminated faster than endocrine cells, and β-cells were lost at a higher rate than α-cells. Interestingly, a specific group of cells that were classified as α-cells but have immature β-cells-like characteristics, were enriched in LTC, revealed the possibility of transdifferentiating of α-cells to β-cells under in vitro culture. Our results suggested that there are intrinsic cellular and molecular mechanisms in pancreatic cells that are associated with their maturity and correlated with their survival ability under unfavorable living conditions.
2025-01-15 | GSE280984 | GEO
Project description:β-cell dysfunction in pancreatic islets is the leading cause of progression to type I diabetes (T1D), but the reason for the loss of β-cell mass in T1D is still not fully understood. Using in vitro long-term cultured (LTC) human islets, we have investigated gene profiling and subtype cells changes in the leftover cells and compared them with residual cells in samples of T1D or T2D. While most of the dysregulated genes were downregulated during LTC, we found there are specific groups of mRNA or miRNA that were upregulated, and they are involved in specific pathways. In general, α-cells and β-cells of LTC-islets have elevated expressions of MAFB and MAFA, respectively. We also found exocrine cells were eliminated faster than endocrine cells, and β-cells were lost at a higher rate than α-cells. Interestingly, a specific group of cells that were classified as α-cells but have immature β-cells-like characteristics, were enriched in LTC, revealed the possibility of transdifferentiating of α-cells to β-cells under in vitro culture. Our results suggested that there are intrinsic cellular and molecular mechanisms in pancreatic cells that are associated with their maturity and correlated with their survival ability under unfavorable living conditions.
2025-01-15 | GSE280940 | GEO
Project description:Transcriptomics and single-cell RNA sequencing analysis of long-term cultured human islets in vitro [miRNA-Seq]
| PRJNA1181523 | ENA
Project description:Transcriptomics and single-cell RNA sequencing analysis of long-term cultured human islets in vitro [scRNA-Seq]
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Project description:RNA sequencing revealed the different transcriptional profiles of long-term cultured and short-term cultured CAR-T cells
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Project description:In this study we show, for the first time, that whole islets can be cultured and significantly expanded long-term in vitro on specific basement membrane (BM) laminin LN-521 which is a normal component of islet BMs. Isolated islets adhere and flatten to form 2-3 cell layer entities with ~10 % of all endocrine cell types proliferating after 2-5 weeks in culture. Time-lapse imaging of freshly isolated islets in suspension labeled with hypoxia and DNA degradation indicators reveals that spherical islets on LN-111 rapidly develop hypoxia and central necrosis, in contrast to islets attached to LN-521, which normal b-cell glucose responsiveness. RNA sequencing analyses reveal that freshly isolated islets express LN-521 binding integrins. Compared to other matrices, islets grown on LN-521 develop a unique expression signature, including expression of specific integrins, involved in cell adhesion and matrix production as well as proliferation. The results reveal specific LN-521-mediated effects into islet cells and suggest that use of in vitro expanded islets may significantly enhance the efficacy of islet transplantation treatment of T1D.
2018-11-09 | GSE89677 | GEO
Project description:Islets are known to respond to changes in ambient glucose. To quantify the transcriptome-wide changes in ambient glucose, we compared transcriptome of islets exposed to low and high glucose. Isolated islets from wild type male mice. Islets from adult males were pooled, cultured overnight in RPMI containing 11 mM glucose. The next day, all islets were starved in RPMI containing 2.8 mM glucose for 2 hours before stimulation with 2.8 mM glucose or 16.8 mM glucose for 12 hours. Islets were lysed in Trizol for RNA isolation and library construction.
2014-09-16 | E-GEOD-58384 | biostudies-arrayexpress
Project description:To evaluate the short-term effect of elastase inhibition using telaprevir or sivelestat or DMSO on human islets
2024-02-13 | GSE189986 | GEO
Project description:M-NM-2-cell identity is determined by tightly regulated transcriptional networks that are modulated by extracellular cues, thereby ensuring M-NM-2-cell adaptation to the organismM-bM-^@M-^Ys insulin demands. We have observed in pancreatic islets that stimulatory glucose concentrations induced a gene profile that was similar to that of freshly isolated islets, indicating that glucose-elicited cues are involved in maintaining M-NM-2-cell identity. Low glucose induces the expression of ubiquitous genes involved in stress responses, nutrient sensing, and organelle biogenesis. By contrast, stimulatory glucose concentrations activate genes with a more restricted expression pattern (M-NM-2- and neuronal- cell identity). Consistently, glucose-induced genes are globally reduced in islets deficient with Hnf1a (MODY3), characterized by a deficient glucose metabolism. Of interest, a cell cycle gene module was the most enriched among the variable genes between intermediate and stimulatory glucose concentrations. Glucose regulation of the islet transcriptome was unexpectedly broadly maintained in islets from aged mice. However, the cell cycle gene module is selectively lost in old islets and the glucose activation of this module is not recovered even in the absence of the cell cycle inhibitor p16. We used microarrays to detail the global programme of gene expression regulated by glucose in young and aged pancreatic islets as well as freshly-isolated islets. Pancreatic islets from young and old mice were isolated and cultured at different glucose concentrations for RNA extraction and hybridization on Affymetrix microarrays. Islets were cultured at 3mM (G3), 5.5mM (G5), 11mM (G11) and 16mM (G16). Freshly-isolated islets (F) were also processed for RNA extraction . We also assessed the dynamic glucose regulation of gene expression at different time-points after an overnight at 3mM (T0): after 1h at 11mM (T1) and after 4h (T4).
2013-07-01 | E-GEOD-42591 | biostudies-arrayexpress