Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation.

Full-term amniotic fluid stem cell (AFSC) is an underexplored reserve of broadly multipotent stem cells with potential applications in cell replacement therapy. One aspect worth exploring is the potential of AFSCs to differentiate into neural lineages. Previously, we have shown that full-term AFSC lines established from term gestation amniotic fluid, known as R3 and R2, differentiated into neural lineage through the monolayer adherent method suggesting their neurogenic potential. The neural commitment of the cells through the formation of multicellular aggregates has never been shown before. Here, we explored the ability of R3 to commit to neural fate via the formation of three-dimensional multicellular aggregates, namely embryoid bodies (EBs) and neurospheres, exhibiting distinct characteristics resembling EBs and neurospheres as obtained from other published pluripotent and neural stem cells (NSCs), respectively. Different cell seeding densities of the cells cultured in their respective induction medium generated two distinct types of aggregates with the appropriate sizes for EBs (300-350 µm) and neurospheres (50-100 µm). The neurospheres expressed a significantly high level of Nestin than EBs. However, EBs stained positive for TUJ1, suggesting the presence of early post-mitotic neurons representing the ectodermal lineage. In contrast, the presence of the NSC population in neurosphere culture was validated with positive expression of Sox1. Notably, dissociated cells from both aggregates differentiated into MAP2-positive neural cells, highlighting the ability of both types of multicellular aggregates to commit to the neural fate. In conclusion, this study highlights the first evidence of neurosphere formation from full-term AFSCs in addition to neural fate commitment via EBs formation. Findings from this study allow researchers to select the suitable approach for neural cell generation and expansion according to research needs.

Caffeine-Supplemented Diet Prevents Fatigue-Like Behavior in Tumor-Bearing Mice.

Caffeine is a widely consumed stimulant, known for its positive effects on physical and mental performance. These effects are potentially beneficial for ameliorating cancer-related fatigue, which affects the quality of life of patients with cancer. This study aimed to determine the anti-fatigue and antitumor effects of caffeine in tumor-bearing mice. BALB/c mice were intravenously injected with C26 colon carcinoma cells and fed with normal or 0.05% caffeine-supplemented diet. Fatigue-like behavior was assessed by running performance using a treadmill test. Lung, blood, liver, muscle, and epididymal adipose tissue samples were collected on day 13 and examined. The antitumor effect of caffeine was assessed using subcutaneous tumor-bearing mice fed with 0.05% caffeine-supplemented diet, and the tumor volume was measured. C26 tumor-bearing mice showed fatigue-like behavior associated with hypoglycemia, depleted liver glycogen and non-esterified fatty acid (NEFA) levels. C26 tumor-bearing mice fed with 0.05% caffeine-supplemented diet showed improved running performance associated with restored NEFA levels. However, exacerbated hypoglycemia and liver glycogen levels after caffeine consumption may be due to tumor-induced catabolic signals, as the tumor volume was not affected. Collectively, caffeine may exert anti-fatigue effects through enhanced lipolysis leading to restored NEFA levels, which can be used as an alternative energy source.

Pax6 mutant cerebral organoids partially recapitulate phenotypes of Pax6 mutant mouse strains.

Cerebral organoids show great promise as tools to unravel the complex mechanisms by which the mammalian brain develops during embryogenesis. We generated mouse cerebral organoids harbouring constitutive or conditional mutations in Pax6, which encodes a transcription factor with multiple important roles in brain development. By comparing the phenotypes of mutant organoids with the well-described phenotypes of Pax6 mutant mouse embryos, we evaluated the extent to which cerebral organoids reproduce phenotypes previously described in vivo. Organoids lacking Pax6 showed multiple phenotypes associated with its activity in mice, including precocious neural differentiation, altered cell cycle and an increase in abventricular mitoses. Neural progenitors in both Pax6 mutant and wild type control organoids cycled more slowly than their in vivo counterparts, but nonetheless we were able to identify clear changes to cell cycle attributable to the absence of Pax6. Our findings support the value of cerebral organoids as tools to explore mechanisms of brain development, complementing the use of mouse models.

Effect of Sucrose on Cisplatin-induced Fatigue-like Behavior in Mice: Comparison With Fructose and Glucose.

Background/Aim: Fatigue is the most common symptom in patients with cancer undergoing radiation therapy or cancer chemotherapy. However, cancer-related fatigue remains undertreated and poorly understood. Materials and Methods: Mice were administered a single dose of cisplatin (10 mg/kg, intraperitoneally) or saline (as a control) and then treated with sucrose, fructose, glucose (each at 500 or 5,000 mg/kg, orally), or saline (control) daily for 4 days. cisplatin-induced fatigue-like behavior was investigated by assessment of running activity on a treadmill. The influence of glucose intake on tumor growth was also examined in Lewis lung carcinoma (LLC)-bearing mice. Results: Administration of sucrose and glucose improved cisplatin-induced fatigue-like behavior in mice, whereas administration of fructose showed only slight antifatigue effects. Although glucose-fed mice showed increased tumor growth, this was balanced out by the powerful cytotoxicity of cisplatin. Conclusion: Sucrose, and especially glucose, may improve patient quality of life during treatment with anticancer agents by preventing fatigue without interfering with the antitumor effects of cisplatin.

Rat full term amniotic fluid harbors highly potent stem cells.

Amniotic fluid stem cells (AFSCs) are commonly isolated from mid-term amniotic fluid (AF) of animals and human collected via an invasive technique, amniocentesis. Alternatively, AFSCs could be collected at full-term. However, it is unclear whether AFSCs are present in the AF at full term. Here, we aimed to isolate and characterize stem cells isolated from AF of full term pregnant rats. Three stem cell lines have been established following immuno-selection against the stem cell marker, c-kit. Two of the new lines expressed multiple markers of pluripotency until more than passage 90. Further, they spontaneously differentiated into derivatives of the three primary germ layers through the formation of good quality embryoid bodies (EBs), and can be directly differentiated into neural lineage. Their strong stemness and potent neurogenic properties highlight the presence of highly potent stem cells in AF of full-term pregnancies, which could serve as a potential source of stem cells for regenerative medicine.