Human platelet lysate efficiency, stability, and optimal heparin concentration required in culture of mammalian cells

BACKGROUND: Fetal bovine serum (FBS) has been used to support the growth and proliferation of mammalian cells for decades. Owing to several risk factors associated with FBS, several trials have been conducted to evaluate substitutes to FBS with the same efficiency and the lower risk issues.METHODS: In this study, human platelet lysate (HPL) derived from activated human platelets was evaluated as an alternative to FBS due to the associated risk factors. To evaluate the efficiency of the preparation process, platelet count was performed before and after activation. The concentrations of several growth factors and proteins were measured to investigate HPL efficiency. HPL stability was studied at regular intervals, and optimal heparin concentration required to prevent gel formation in various media was determined. The biological activity of HPL and FBS was compared by evaluating the growth performance of Vero and Hep-2 cell lines.RESULTS: Result of platelet count assay revealed the efficiency of HPL preparation process. Growth factor concentrations in HPL were significantly higher than those in FBS, while the protein content of HPL was lower than that of FBS. Stability study data showed that the prepared HPL was stable for up to 15 months at −20℃. Ideal heparin concentration to be used in different media was dependent on calcium concentration. Results of cell viability assay showed that HPL was superior to FBS in supporting the growth and proliferation of Vero and Hep-2 cells.CONCLUSION: The HPL prepared by the mechanical activation of platelets may serve as an efficient alternative to FBS in cell culture process.

Renal ischemia -reperfusion injury: contribution of nitric oxide and reactive oxygen species

The present study was undertaken to examine the effect of ischemia/reperfusion injury to the kidney on basal NO production and to define whether the antioxidants or NO precursor can modulate NO production in this model or not. Induction of renal ischemia for 60 minutes followed by 15 minutes reperfusion in adult male Wistar rats resulted into impairment of NO production as indicated by reduced serum nitrite/nitrate levels, increase in lipid peroxidation measured as malondialdehyde [MDA] and depletion of reduced glutathione [GSH] and catalase contents in rat kidney. NO precursor, L-arginine [L-ARG] as well as hydroxyl radical scavengers, dimethylthiourea [DMTU] and an iron chelator, deferoxamine [DFO] increased serum NO level and renal GSH and catalase levels and reduced renal MDA. Conversely, infusion of iron complex EDTA-FeCl3 10 minutes before ischemia exacerbated the changes in the parameters studied was induced by ischemia/reperfusion

Effect of curcumin on cadmium- induced hepatic toxicity in rats

The protective effect of spice principle, curcumin on cadmium [Cd] induced hepatotoxicity in rats were studied. Male albino rats injected i.v. with 4 mg CdCl[2]/Kg showed hepatic damage as measured by an increase in liver lipid peroxide product as malondialdehyde [MDA], glutathione peroxidase [GProx] and glutathione reductase [GRed]. On the other hand, hepatic superoxide dismutase [SOD] and reduced glutathione [GSH] content decreased significantly. The increase in liver MDA concomitant with elevated serum enzymes, alanine aminotransferase [ALAT], aspartate aminotransferase [ASAT] and lactate dehydrogenase [LDH]. Pre-administration of curcumin [30 mg/kg body wt.] for 7 days succeeded to ameliorate hepatic MDA. SOD and GSH in Cd-injected rats. The levels of GProx and GRed being normalized. Curcumin also lowered significantly serum ALAT, ASAT and LDH levels. These results elucidate the potent antioxidant protective action of curcumin against cadmium-induced hepatic damage as monitered by lipid peroxide and antioxidant enzymes

Role of hypolipidemic doses of nicotinic acid on glucose tolerance and severity of diabetes in male rats

The present work was devoted to study the effect of varying doses of nicotinic acid on glucose tolerance, and its role in modulating the severity of STZ- diabetes in male rats. Three oral nicotinic acid doses of, 31 5, 63.5 and 126.0 mg/kg b. w. respectively, comparable to half, equal and two folds rodent equivalent of 600 mg [daily human hypolipidemnic dose], were administered to three equal sized groups of male albino rats, prior to oral glucose load of 1.0 g/kg b.w. Serial post loading glucose determinations, revealed marked increases in glucose tolerance, in parallel with the rise in nicotinic acid dosage, as verified by lower magnitudes of post-loading hyperglycaema, compaired to parallel non-medicated Controls. Moreover, the results of serum glucose 'FSG estimates in groups of STZ- diabetic rats, receiving daily repeated oral nicotinic acid medication in doses of 63.0 and 126.0 mg/kg. b.w. respectively, yielded evidence of very highly significant magnitudes of antagonistic attenuation of STZ- hyperglycaemia, coupled with marked increase in serum insulin activity, progressively accentuated with longer duration of medication