- Transcriptome analysis of the. - Background: Bone marrow mesenchymal stem cells are a potential resource for the clinical therapy of certain diseases. - Canine, as a companion animal, living in the same space with human, is an ideal new model for human diseases research. - Because of the high prevalence of diabetes, alternative transplantation islets resource (i.e. - insulin producing cells) for diabetes treatment will be in urgent need, which makes our research on the transdifferentiation of Bone marrow mesenchymal stem cells into insulin producing cells become more important.. - stage 2 ” and “ stage 3. - A total of 11,530 differentially expressed transcripts were revealed in the profiling data. - Conclusion: In conclusion, to the best of our knowledge, this is the first survey of the transdifferentiation of canine BMSCs into insulin-producing cells according with the timeline using next-generation sequencing technology. - Stem cells in particular for BMSCs have been used for decades for the treatment of many diseases. - producing cells [4]. - The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. - The College of Veterinary Medicine of the Northwest Agriculture and Forestry University, No.3 Taicheng Road, Yangling 712100, Shaanxi, P. - However, all of the available reprogramming methods are limited by the low efficiency of transformation which may lead to an insufficiency of pancreatic islets for transplantation surgery. - This problem is associ- ated with our limited understanding of the transdifferen- tiation mechanisms.. - Induction and characterization of islet-like spheroids At the nonadherent stage, BMSCs formed many spheroids floating in the medium (Fig. - a 24 h after the separation, only blood cells could be seen in the dish. - representing 67.60% of the total on average.. - 2c), while there were 201 upregulated genes and 213 down- regulated ones for the comparison between “stage 1” and. - 2d), 232 and 132 for “stage 2” and “stage. - 2e), and 2331 and 2758 for “islets” vs “stage 3,”. - All of the DEGs, including upregulated and downregu- lated transcripts, were annotated to signaling pathways related to the GO terms. - stage 1” DEGs were mapped to 230 KEGG pathways, while the “stage 1 vs. - stage 2,” “stage 2 vs. - 3 Top 20 KEGG signaling pathways of known transcripts, the size of bubbles represented for mapped gene numbers of the item, and the color was measured by the Qvalue. - a KEGG analysis of “ BMSCs vs stage 1. - b KEGG analysis of “ stage 1 vs stage 2. - c KEGG analysis of “ stage 2 vs stage 3. - d KEGG analysis of “ islets vs stage 3. - The GO analysis of “BMSCs vs. - For “stage 1 vs. - stage 2,” the DEGs were mainly anno- tated to the categories of cellular protein modification process, protein modification process, and phosphate- containing compound metabolic process for BP;. - Then, for the. - They were chosen from two datasets, GSE20113 and GSE52063, which belong to the same platform, GPL3738. - Because of the different backgrounds of the samples, the normalization of these data was performed before enrichment analysis using R packages limma and gplots (Fig. - There were 1431 DEGs between those two series, 771 downregulated and 660 upregulated, showed in the heatmap (Fig. - In terms of the GO results, chaperone-mediated protein folding and cellular zinc ion. - 5 Normalization and enrichment analysis of GEO data. - a The status of the raw data from GEO database showed in box-plot. - and ion channel activity and oxido- reductase activity for MF (Additional file 4) are essential processes of the pancreas. - All these results are analogous to the findings of our samples from the transition of. - After the induction proced- ure, it was showed that the expression level of PDX1, NGN3 and NKX2.2 were upregulated in the overexpres- sion groups of VIP and RPS6KA6 (Fig. - 6 PPI network analysis of known transcripts. - Screening for DEGs between different timepoints was performed, and refer- ring to pancreatic development, we could tell what state the cells were in during the induction, enabling adjust- ment of the induction procedure to achieve optimal con- ditions. - However, surprisingly, besides our team, no other groups have focused on the timeline- based analysis of the whole transition process of canine BMSCs, and with transcriptome profiling, the data has been uploaded on SRA database (https://www.ncbi.nlm.. - Combined with the sequencing data, we found that all of the transcriptome profiling data and GEO data pro- vided the same findings. - The increased GO term binding activity including protein binding and ion bind- ing reflected certain characteristics of the pancreas, gen- erally related to vigorous secretory activity and calcium exchange [49]. - 7 a The verification of the 20 key genes by qRT-PCR. - Interestingly, SSTR2 may involve in the rhythmic glucagon and insulin secre- tion [60]. - Furthermore, these genes were participating in the same pathway like MAPK sig- naling pathway [61], Rap1 signaling pathway and Ras signaling pathway, and it could be a lead for the next- step mechanism research of the induction process to obtain high quality islet-like clusters.. - In this study, we have obtained an overview of the path- ways involved in the regulation of BMSC transdifferen- tiation at different timepoints for the first time, and it makes a progress for learning the mechanism. - We supposed that fine-tuning of these genes might contribute to transdifferentiation of BMSCs into IPCs and make an advance to the induction procedure.. - The protocol used in this study involved three stages, while the BMSCs were placed in a suspended state to form spheroids, which mimicked the characteristics of islets in the pancreas. - The dishes used in the protocol were all treated with 2-hydroxyethyl methacrylate (Sigma-Aldrich). - Enrichment analysis of DEGs and PPI analysis. - Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was also adopted to define the func- tions of the DEGs. - The significance formula was the same as that in the GO analysis, and it was presented using the magnitude of –log10 (Q-value) (Q-value was derived from multiple hypothesis testing of the P-value). - Mean- while, RF (rich factor) was defined as the ratio of DEGs relative to the overall annotated transcripts enriched in the same pathway.. - Canine data were included in the database, so we constructed the net- works by extracting the target gene list from the data- base. - Because these data were from the same platform, after normalization of the microarray data, we used R (https://www.r-project.org/) to acquire the DEGs, and analyzed these data using the DAVID database (https://. - A summary of the quality of output data. - The number of reads multiplied by the length of the sequence is converted to G. - Q30 ” represented the percentage of bases with Phred value greater than 20 and 30 in the total base. - The number of reads that can be mapped to the genome. - The reads that can map to the sense strand of genome. - The reads that can map to the antisense strand of genome. - The number of the intact Uniquely mapped reads which. - The number of the segmented Uniquely mapped reads which map to exons. - Additional file 3 KEGG analysis of GEO data using DAVID. - Top 20 items of GO analysis of GEO data. - BMSCs: Bone marrow mesenchymal stem cells. - This work was supported by the National Natural Science Foundation of China (project number in the design of study, data collection, analysis .and writing of the manuscript.. - The application named “ The use of two Chinese rural dogs for the separation of BMSCs ” were approved by the Animal Ethics Committee of Northwest Agriculture and Forest University (Animal Welfare Assurance no.. - The informed consent was verbally permitted by the leader of institution Yihua Zhang, because there was no need to provide additional written information under the ap- proval of the committee.. - Clinical use of bone marrow, bone marrow concentrate, and expanded bone marrow mesenchymal stem cells in cartilage disease. - Stem Cells Dev. - Advances of mesenchymal stem cells derived from bone marrow and dental tissue in craniofacial tissue engineering. - Bone marrow mesenchymal stem cells (BMSCs) improved functional recovery of spinal cord injury partly by promoting axonal regeneration. - Stem cell-based treatments for type 1 diabetes mellitus: bone marrow, embryonic, hepatic, pancreatic and induced pluripotent stem cells. - Genome-wide analysis reveals changes in long noncoding RNAs in the differentiation of canine BMSCs into insulin-producing cells. - Mesenchymal stem cells. - Stem cells in the treatment of diabetes mellitus - focus on mesenchymal stem cells. - Stem Cells Int. - in mice with an inducible adipocyte-specific deletion of the insulin receptor. - Differentiation of human embryonic stem cells into insulin-producing clusters. - Stem cells (Dayton, Ohio). - Under a nonadherent state, bone marrow mesenchymal stem cells can be efficiently induced into functional islet-like cell clusters to normalize hyperglycemia in mice: a control study. - Isolation and culture of bone marrow Mesenchymal stem cells from human fetus and its biological properties. - Overexpression of ERBB4 rejuvenates aged mesenchymal stem cells and enhances angiogenesis via PI3K/AKT and MAPK/ERK pathways. - Bone marrow-derived Mesenchymal stem cells protect islet grafts against endoplasmic reticulum stress-induced apoptosis during the early stage after transplantation. - Stem Cells. - A modified method of insulin producing cells' generation from bone marrow-derived mesenchymal stem cells. - Mesenchymal stem cells in the treatment of type 1 diabetes mellitus. - T UG: in vitro generated Mesenchymal stem cells:. - Somatostatin and insulin mediate glucose- inhibited glucagon secretion in the pancreatic α -cell by lowering cAMP. - Curcumin-mediated bone marrow mesenchymal stem cell sheets create a favorable immune. - Stem Cells Transl Med. - Gap junctional intercellular communication in adipose-derived stromal/stem cells is cell density-dependent and positively impacts adipogenic differentiation. - Vasoactive intestinal peptide stimulates bone marrow- Mesenchymal stem cells Osteogenesis differentiation by activating Wnt/ β - catenin signaling pathway and promotes rat skull defect repair. - Transplantation of amniotic scaffold seeded mesenchymal stem cells and/or endothelial progenitor cells from bone marrow to efficiently repair 3-cm circumferential urethral defect in model dogs. - Immortalization of canine adipose- derived mesenchymal stem cells and their seminiferous tubule transplantation
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