Generation of a human induced pluripotent stem cell line overexpressing CCL22 with islet cells differentiation potential.

CCL22 is a macrophage-derived immunosuppressive chemokine that recruits regulatory T cells through the CCL22:CCR4 axis, playing an important role in homeostatic and inflammatory responses. A CCL22-overexpressing human induced pluripotent stem cell line (CNNDi001-A-2) was generated by lentiviral transduction to further study the function of CCL22. The cell line was confirmed to have normal proliferation and pluripotency and could be further differentiated into islet cells for cell replacement therapy in diabetes.

Inhibition of the Rho/ROCK pathway promotes the expression of developmental and migration-related genes in astrocytes exposed to alcohol.

Astrocytes are an important regulator of alcohol dependence. Furthermore, the downregulation of Rho-associated coiled coil-containing protein kinase 2 (ROCK2) attenuates alcohol-induced inflammation and oxidative stress in astrocytes. On the basis of these findings, we examined the effects of alcohol and a Rho/RACK kinases inhibitor on astrocyte function and investigated their effects on mRNA expression to further explore the protective mechanisms of a Rho/RACK kinases inhibitor in astrocytes after alcohol exposure. CTX TNA2 astrocytes were cultured with alcohol and Rho/RACK kinases inhibitor intervention before undergoing transcriptome sequencing, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and wound healing assays. Alcohol exposure modulated cell morphology and inhibited astrocyte migration, whereas Fasudil improved cell morphology and promoted astrocyte migration after alcohol exposure. Transcriptome sequencing results indicated that alcohol exposure modulates the expression of genes involved in astrocyte development. Fasudil reversed the effects of alcohol exposure on the astrocyte developmental process. Four genes related to the developmental process and migration - Ccl2, Postn, Itga8, and Serpine1 - with the highest protein-protein interaction correlations (node degree >7) were selected for verification by qRT-PCR, and the results were consistent with those of the sequencing and wound healing assays. Our results suggest that the Rho/ROCK pathway is essential for alcohol to be able to interfere with astrocyte development and migration gene expression. The Rho/ROCK pathway inhibitor Fasudil reversed the adverse effects of alcohol exposure on astrocytes and may have clinical applications.

Effects of Notch-1 signalling pathway on differentiation of marrow mesenchymal stem cells into neurons in vitro.

In this study, the putative involvement of the Notch-1 signaling pathway in the neuronal differentiation of bone marrow mesenchymal stem cells (BMSCs) was investigated through RNA interference (RNAi). We found that mNotch-1shRNA could efficiently block expression of Notch-1 in BMSCs. After induction, immunohistochemistry, reverse transcriptase-PCR and Western blot analyses, all indicated that the expression of neuron-specific markers such as neuron-specific enolase and neurofilament 200 was much higher in mNotch-1shRNA BMSCs than that in control groups, whereas the expression of glial fibrillary acidic protein was lower in mNotch-1shRNA BMSCs. The percentage of apoptotic cells in mNotch-1shRNA BMSCs, however, was significantly higher than that in control groups. These data indicate that Notch signaling plays a role in the differentiation of BMSCs into neurons in vitro. Pharmacological or genetic interference with Notch signaling may provide a novel method to obtain neurons for therapeutic use.