Transcriptome analysis of the transdifferentiation of canine BMSCs into insulin producing cells

Abstract

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. Result In this study, we completed the transdifferentiation process and achieved the transcriptome profiling of five samples with two biological duplicates, namely, “BMSCs”, “islets”, “stage 1”, “stage 2” and “stage 3”, and the latter three samples were achieved on the second, fifth and eighth day of induction. A total of 11,530 differentially expressed transcripts were revealed in the profiling data. The enrichment analysis of differentially expressed genes revealed several signaling pathways that are essential for regulating proliferation and transdifferentiation, including focal adhesion, ECM-receptor interaction, tight junction, protein digestion and absorption, and the Rap1 signaling pathway. Meanwhile, the obtained protein–protein interaction network and functional identification indicating involvement of three genes, SSTR2, RPS6KA6, and VIP could act as a foundation for further research. 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. The three key genes we pick out may regulate decisive genes during the development of transdifferentiation of insulin producing cells.

Novel Functional Genes Involved in Transdifferentiation of Canine ADMSCs Into Insulin-Producing Cells, as Determined by Absolute Quantitative Transcriptome Sequencing Analysis

“…In KEGG enrichment analysis, DEGs are mainly concentrated in tight junction, protein digestion and absorption, pancreatic secretion, focal adhesion, ECM-receptor interaction, Rap1 signaling pathway, and cell cycle, etc. In GO enrichment analysis, DEGs are mainly concentrated in the categories of nucleus, extracellular region, intracellular membrane-bound organelle, the regulation of transcription, regulation of RNA biosynthetic process, carbohydrate metabolic process, single-organism carbohydrate metabolic process and small GTPase-mediated signal transduction, et al Sstr2 , Rps6ka6 , and Vip they pick up may regulate decisive genes during the development of transdifferentiation of insulin producing cells ( Wang et al, 2021 ). In this study, Absolute Quantitative Transcriptome Sequencing was used to detect the changes of genes and metabolic pathways during the transdifferentiation of canine ADMSCs into IPCs in vitro for the first time.…”

Short-term transplantation effect of a tissue-engineered meniscus constructed using drilled allogeneic acellular meniscus and BMSCs

“…The identified 4th passage canine BMSCs frozen in the laboratory ( 13 , 14 ) were amplified for 2 passages using complete α-MEM (α-MEM culture medium containing 10% fetal bovine serum, 1% 100 × Penicillin Streptomycin Solution and 0.25% Anti-Myc Mycoplasma scavenging reagent) in 60 mm petri dishes. The 6th passage BMSCs were prepared into 3 × 10 5 /mL cell suspensions, and 500 μL of the cell suspensions were inoculated (uniform drop cloth) on the drilling meniscus ( n = 3) and the control meniscus ( n = 3).…”