Background Insulin producing cells generated by liver cell transdifferentiation, could serve as an attractive source for regenerative medicine. The present study assesses the relationship between DNA methylation pTFs induced liver to pancreas transdifferentiation. Results The transdifferentiation process is associated with DNA demethylation, mainly at gene regulatory sites, and with increased expression of these genes. Active inhibition of DNA methylation promotes the pancreatic transcription factor-induced transdifferentiation process, supporting a causal role for DNA demethylation in this process. Conclusions Transdifferentiation is associated with global DNA hypomethylation, and with increased expression of specific demethylated genes. A combination of epigenetic modulators may be used to increase chromatin accessibility of the pancreatic transcription factors, thus promoting the efficiency of the developmental process.

Genetic manipulations can ameliorate the aging process and extend the lifespan of model organisms. The aim of this research was to identify novel genetic interventions that promote both lifespan and healthspan, by combining the effects of multiple longevity-associated gene inactivations in C. elegans. For this, the individual and combined effects of the odr-3 mutation and of ife-2 and cku-70 knock-downs were studied, both in the wild type and daf-16 mutant backgrounds. We found that besides increasing the lifespan of wild type animals, the knock-down of ife-2 (starting at L4) also extends the lifespan and healthspan of long-lived odr-3 mutants. In the daf-16 background, ife-2 and odr-3 impairment exert opposing effects individually, while the daf-16; odr-3; ife-2 deficient animals show a similar lifespan and healthspan as daf-16, suggesting that the odr-3 and ife-2 effector outcomes converge downstream of DAF-16. By contrast, cku-70 knock-down did not extend the lifespan of single or double odr-3; ife-2 inactivated animals, and was slightly deleterious to healthspan. In conclusion, we report that impairment of odr-3 and ife-2 increases lifespan and healthspan in an additive and synergistic manner, respectively, and that this result is not improved by further knocking-down cku-70.

Autologous cells replacement therapy by liver to pancreas transdifferentiation (TD) allows diabetic patients to be also the donors of their own therapeutic tissue. Aim: To analyze whether the efficiency of the process is affected by liver donors' heterogeneity with regard to age, gender and the metabolic state. Materials & methods: TD of liver cells derived from nondiabetic and diabetic donors at different ages was characterized at molecular and cellular levels, in vitro. Results: Neither liver cells proliferation nor the propagated cells TD efficiency directly correlate with the age (3-60 years), gender or the metabolic state of the donors. Conclusion: Human liver cells derived from a wide array of ages and metabolic states can be used for autologous cells therapies for diabetics.

Background: Liver cells represent an attractive source of cells for autologous regenerative medicine. The present study assesses the liver cells' stability during in vitro expansion, as a prerequisite for therapeutic use. Results: The human liver cell cultures in this study were propagated efficiently in vitro for at least 12 passages. No significant changes in morphology, intracellular ultrastructures and characteristic markers expression were found during in vitro expansion of cells from all analyzed donors. However, expanded cells derived from male donors of >60 years old, lost the Y chromosome. Conclusion: Liver-derived cell cultures adopt a proliferative, stable mesenchymal phenotype, through an epithelial to mesenchymal transition process. The molecular and phenotypic changes of the cells during propagation are uniform, despite the heterogeneity of the different donors. Loss of Y chromosome occurs after cells' propagation in elder male donors.

Transdifferentiation is the direct reprogramming of adult cells into alternate cell types with different function. Liver to pancreas transdifferentiation (TD) induced by ectopic expression of pancreatic transcription factors (pTFs) was first described by our group both in vivo (1) and in human liver cells in vitro (2). Aim: Disclose the mechanism that mediate the developmental reprogramming process of adult human liver cells into endocrine pancreatic cells. Determine the developmental barriers that restrict this process efficiency and suggest modalities to increase the process efficiency. Finally, the clinical and industrial translation of adult cells reprogramming will be discussed.

AIM:Glucotoxicity obstructs pancreatic differentiation from adult stem cells. The aim was to develop a novel protocol for differentiation of dental pulp stem cells (DPSCs) into pancreatic β cells and determine the effect of H2S on glucotoxicity.MATERIALS & METHODS:DPSCs were differentiated with media containing 5.5 or 25.0 mM glucose, exposed to 1 ng/ml H2S. Glucotoxicity, expression of β-cell markers, INS, PDX1 and GLUT2, and PI3K/AKT pathway were assessed.RESULTS:H2S exposure increased insulin and C-peptide, and protected DPSC-derived pancreatic β-like cells from glucotoxicity and upregulated INS, PDX1 and GLUT2, and genes of PI3K/AKT pathway.CONCLUSION:H2S improved effects of glucotoxicity on β-like cells via PI3K/AKT pathway. The protocol for pancreatic β-cell differentiation might have applications in regenerative medicine rather than swine pancreas transplantation.

Cell size is, alongside sex and depot of origin, the key determining factor of adipocyte function. Ther e is large variation in this parameter between the di fferent compartments of this adipose organ, owing to differences in cell turn-over, mechanisms of adi pose tissue expansion and a plethora of other physiological causes. The current study aims to ass ess the differences in size and number between adipocytes from two of the most metabolically signi ficant body fat depots, the visceral omental adipose tissue and the abdominal subcutaneous adipo se tissue. Paired samples of adipose tissue were obtained from a group of 15 surgical patients (11 w omen, 4 men) and were subjected to cytomorphometric analysis. We found that mean adipo cyte diameter was significantly larger for abdominal subcutaneous adipocytes than omental adip ocytes (184.9 ± 9.75 μm, compared with 155.96 ± 7.23 μm, respectively; p<0.05) by roughly 15.65%, and that minimum adipocyte diameter was significantly larger for abdominal subcutaneous adipocytes than omental adipocytes (93.76 ± 8.64 μm, compared with 69.05 ± 5.3 μm, respectively ; p<0.05) by roughly 26.35%. No significant correlations were found for maximum adipocyte diame ter (abdominal subcutaneous 272.92 ± 10.24 μm, omental 245.9 ± 13.36 μm; p>0,5) and number of adipocytes per microscopic field (abdominal subcutaneous 335.33 ± 41.37, omental 432.67 ± 56.0 9; p>0.5). The results were consistent with previous findings in scientific literature and with t e view that the two compartments play different metabolic roles and undergo expansion through diffe rent mechanisms.