Proline-based solution maintains cell viability and stemness of canine adipose-derived mesenchymal stem cells after hypothermic storage.

Transportation of mesenchymal stem cells (MSCs) under hypothermic conditions in 0.9% normal saline solution (NSS) might increase cell death and alter the stemness of MSCs. The present study aimed to evaluate the effect of proline-based solution (PL-BS) on cell viability and the stemness of newly established canine adipose-derived mesenchymal stem cells (cAD-MSCs) under hypothermic conditions. Characterized cAD-MSCs were stored in 1, 10, and 100 mM PL-BS or NSS at 4°C for 6, 9, and 12 hours prior to an evaluation. The results demonstrated that storage in 1 mM PL-BS for 6 hours decreased cell apoptosis and proliferation ability, but improved cell viability and mitochondrial membrane potential. cAD-MSCs maintained their high expression of CD44 and CD90, but had a low expression of CD34 and MHC class II. Trilineage differentiation ability of cAD-MSCs was not affected by storage in 1 mM PL-BS. Gene expression analysis demonstrated that immunomodulatory genes, including IDO, HGF, PGE-2, and IL-6, were upregulated in cAD-MSCs stored in 1 mM PL-BS. In conclusion, PL-BS can be effectively applied for storing cAD-MSCs under hypothermic conditions. These findings provide a new solution for effective handling of cAD-MSCs which might be promising for clinical applications.

Melatonin attenuates dimethyl sulfoxide- and Zika virus-induced degeneration of porcine induced neural stem cells.

Domestic pigs have become increasingly popular as a model for human diseases such as neurological diseases. Drug discovery platforms have increasingly been used to identify novel compounds that combat neurodegeneration. Currently, bioactive molecules such as melatonin have been demonstrated to offer a neuroprotective effect in several studies. However, a neurodegenerative platform to study novel compounds in a porcine model has not been fully established. In this study, characterized porcine induced neural stem cells (iNSCs) were used for evaluation of the protective effect of melatonin against chemical and pathogenic stimulation. First, the effects of different concentrations of melatonin on the proliferation of porcine iNSCs were studied. Second, porcine iNSCs were treated with the appropriate concentration of melatonin prior to induced degeneration with dimethyl sulfoxide or Zika virus (ZIKV). The results demonstrated that the percentages of Ki67 expression in porcine iNSCs cultured in 0.1, 1, and 10 nM melatonin were not significantly different from that in the control groups. Melatonin at 1 nM protected porcine iNSCs from DMSO-induced degeneration, as confirmed by a dead cell exclusion assay and mitochondrial membrane potential (ΔΨm) analysis. In addition, pretreatment with melatonin reduced the percentage of dead porcine iNSCs after ZIKV infection. Melatonin increased the ΔΨm, resulting in a decrease in cell degeneration. However, pretreatment with melatonin was unable to suppress ZIKV replication in porcine iNSCs. In conclusion, the present study demonstrated the anti-degenerative effect of melatonin against DMSO- and ZIKV-induced degeneration in porcine iNSCs.

Effect of Curcumin on Collagen Deposition and Tissue Reorganization in Liver of Diabetic Rats.

Objective: To localize and characterize type I and type IV collagens in recovery livers after curcumin supplementation in streptozotocin-induced diabetic rats. Material and Method: Induced diabetic rats were performed by streptozotocin injection (60 mg/kg BW). Male rats were organized into three groups, control rat (C), diabetic rat (DM) and diabetic rat supplemented with curcumin (DMC) (200 mg/kg BW). At 8 weeks, animals were sacrificed. The localization and characterization of type I and type IV collagens in liver's cell and tissues were compared among C, DM and DMC groups by Sirius red and Immunohistochemical techniques, respectively. Results: Type I and type IV collagens might be the key mediators of liver tissue healing associated with various disorders, especially with inflammation and reorganization processes. Concerning diabetic experiments, increased type I collagen was intensively recognized at subendothelial area of central veins whereas weakly demonstrated at periportal triad and perisinusoidal areas. Conversely, the high intensity of distribution of type IV collagen was strongly revealed at periportal triad and perisinusoidal areas while the intensity was faintly presented at central veins. In addition, accumulation of type IV collagen also revealed perisinusoidal basement membrane which was characteristic of capillarization of sinusoids. However, the localization of type I and type IV collagens was reduced after curcumin supplement in DMC rats compared with DM rats, implying that the liver tissue reorganization has been developed forwards to normal morphology. Moreover, type I and type IV collagen might distinctively accomplish the liver tissue reorganization by different means of area-based characterization. Conclusion: The potential beneficial effect of curcumin has been exhibited the tissue reorganization of diabetic liver tissues. The efficiency and achievement of curcumin might be applied to be an alternative therapeutic agent in diabetic hepatic pathology.