Cryopreservation effects on Wharton's Jelly Stem Cells proteome.

Cryopreservation is the only method for long-term storage of viable cells and tissues used for cellular therapy, stem cell transplantation and/or tissue engineering. However, the freeze-thaw process strongly contributes to cell and tissue damage through several mechanisms, including oxidative stress, cell injury from intracellular ice formation and altered physical cellular properties. Our previous proteomics investigation was carried out on Wharton's Jelly Stem Cells (WJSCs) having similar properties to adult mesenchymal stem cells and thus representing a rich source of primitive cells to be potentially used in regenerative medicine. The aim of the present work was to investigate molecular changes that occur in WJSCs proteome in different experimental conditions: fresh primary cell culture and frozen cell. To analyze changes in protein expression of WJSCs undergoing different culturing procedures, we performed a comparative proteomic analysis (2DE followed by MALDI-TOF MS/MS nanoESI-Q-TOF MS coupled with nanoLC) between WJSCs from fresh and frozen cell culturing, respectively. Frozen WJSCs showed qualitative and quantitative changes compared to cells from fresh preparation, expressing proteins involved in replication, cellular defence mechanism and metabolism, that could ensure freeze-thaw survival. The results of this study could play a key role in elucidating possible mechanisms related to maintaining active proliferation and maximal cellular plasticity and thus making the use of WJSCs in cell therapy safe following bio-banking.

Influence of cigarette smoking on vitamin E, vitamin A, beta-carotene and lycopene concentrations in human pre-ovulatory follicular fluid.

Increasing evidence suggests that fat soluble vitamins and micronutrients have the potential for local modulation of follicular development. Cigarette smoking has been associated with accelerated follicular depletion and derangement of reproductive functions. The present study was initiated to investigate the impact of cigarette smoking on follicular and plasma concentrations of vitamin A, vitamin E, lycopene and beta-carotene. Samples were collected from 17 smokers and 43 non-smoking women undergoing assisted reproduction techniques. Assays were carried out by a reverse-phase high-pressure liquid chromatography (HPLC) method. Smokers had significantly (P < 0.05) lower levels of follicular fluid beta-carotene in comparison to non-smokers (0.02 +/- 0.02 vs. 0.09 +/- 0.02, respectively). No other significant influences on follicular and plasma concentrations were noted. Smokers showed a significantly (P < 0.05) lower fertilization rate in comparison to non-smokers, (55.9 % vs. 71.5 % , respectively). It is postulated that follicular depletion of the antioxidant beta-carotene occurs in response to oxidative stress imposed by cigarette smoke.