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Furthermore, the cells have a very low transfection efficiency, batch-to-batch variation and have to be closely monitored across passages to ensure their beta cells characteristics. It is thus not possible to expand a culture from a single cell, which precludes the generation of a modified clonal cell line. The application in EndoC-βH1, however, is not straightforward, as their proliferation rate is low and they are very sensitive to seeding densities. This genome editing tool has revolutionised genetic manipulations by being an easily programmable RNA-guided endonucleaseģ9. Robust protocols for generating gene KO using CRISPR/Cas9 in EndoC-βH1 studies have not yet been described. Further independent investigations have also demonstrated their suitability for both high-throughput screening Multiomic profiling in EndoC-βH1 cells including epigenomic and transcriptomic maps largely recapitulate primary human islets signatures and along with their similar electrophysiological properties, EndoC-βH1 are therefore a representative model of human beta cells and physiological insulin secretionģ0. Insulin content is a magnitude lower than in primary human beta cells, but secreted insulin as percentage of content and the stimulation index are in the same range as for pancreatic isletsĢ7. The isolated and passaged cells were able to secrete insulin in response to different glucose and secretagogues stimulation, expressed key beta cell markers and were negative for other pancreatic cell markers like glucagonĢ6. Between each transduction, the cells were transplanted into SCID mice to expand and form insulinomas. To generate this cell line, fetal pancreatic buds were transduced with oncogene simian virus 40 large tumour antigen (SV40LT) and human telomerase reverse transcriptase (hTERT). In vitro and provided a valuable alternative to rodent beta cell linesĢ6. In 2011, Scharfmann and colleagues released the EndoC-βH1 cell line, a human beta cell line which opened up the possibility of studying human beta cell physiology and pathology Other distinguishing factors are the proliferative capacity and ion channel composition of rodent cells compared to human beta cells INS2, which is in contrast to human beta cells, which only have one insulin geneĢ0. Furthermore, rodent beta cells express two insulin genes, SLC2A3, leading to distinct glucose uptake dynamicsġ9. SLC2A2, whereas human beta cells mainly utilise The principal glucose transporter in rodent beta cells is
Differences in key components of the glucose-stimulated insulin secretion (GSIS) pathway emphasise unique functional signaturesġ6. PPARG, some of which are involved in Type 1 (T1D) and T2D pathologyġ5. However, the studies also highlighted a substantial number of uniquely expressed genes in either species and significantly differentially expressed genes such as Transcriptomic analysis in purified beta cells from mice and human demonstrated a set of common core beta cell genesġ4. Rodent islets on the other hand have a higher vascular density and are made up of a distinct beta cell core and non-beta cell mantleġ2. Human pancreatic islets have a substantially different architecture than rodent islets as they have fewer beta cells, a mixed cell distribution throughout the whole islet, with alpha, beta and delta cells being adjacent to each other and alpha cells clustering around blood vesselsġ1. Although they share many similarities with human beta cells, there are also fundamental structural, transcriptional and functional differences, aside from having a distinctive genetic background.
Meanwhile, rodent beta cell lines have provided valuable insights into beta cell function and pathophysiologyĩ. The vast majority of these exert their impact on T2D-risk through the pancreatic beta cell and therefore authentic human beta cell models are essential for functional follow-up studiesĪ lack of a stable and functional human beta cell line, restricted access to human cadaveric pancreatic islets and functional limitations of induced pluripotent stem cells (iPSC)-derived beta-like cells have long been a challenge in understanding beta cell biology. Genome-wide association studies (GWAS) have so far identified more than 240 loci which are robustly associated with disease riskĥ. Type 2 diabetes (T2D) affects around 400 million people worldwide and is a complex disease with genetic and non-genetic risk factorsġ.