ABSTRACT
BACKGROUND: Menstrual blood-derived stem cells (MenSCs) are a novel source of stem cells that can be easily isolated non-invasively from female volunteered donor without ethical consideration. These mesenchymal-like stem cells have high rate of proliferation and possess multi lineage differentiation potency. This study was undertaken to isolate the MenSCs and assess their potential in differentiation into epidermal lineage. METHODS: About 5-10 ml of menstrual blood (MB) was collected using sterile Diva cups inserted into vagina during menstruation from volunteered healthy fertile women aged between 22-30 years. MB was transferred into Falcon tubes containing phosphate buffered saline (PBS) without Ca2(+) or Mg2(+) supplemented with 2.5 µg/ml fungizone, 100 µg/mL streptomycin, 100 U/mL penicillin and 0.5 mM EDTA. Mononuclear cells were separated using Ficoll-Hypaque density gradient centrifugation and washed out in PBS. The cell pellet was suspended in DMEM-F12 medium supplemented with 10% FBS and cultured in tissue culture plates. The isolated cells were co-cultured with keratinocytes derived from the foreskin of healthy newborn male aged 2-10 months who was a candidate for circumcision for differentiation into epidermal lineage. RESULTS: The isolated MenSCs were adhered to the plate and exhibited spindle-shaped morphology. Flow cytometric analysis revealed the expression of mesenchymal markers of CD10, CD29, CD73, and CD105 and lack of hematopoietic stem cells markers. An early success in derivation of epidermal lineage from MenSCs was visible. CONCLUSION: The MenSCs are a real source to design differentiation to epidermal cells that can be used non-invasively in various dermatological lesions and diseases.
ABSTRACT
Rapid Infiltration Basin Systems (RIBS) are used for disposing reclaimed wastewater into soil to achieve additional treatment before it recharges groundwater. Effluent from most new sequenced batch reactor wastewater treatment plants is completely nitrified, and denitrification (DNF) is the main reaction for N removal. To characterize effects of complex surface and subsurface flow patterns caused by non-uniform flooding on DNF, a coupled overland flow-vadose zone model is implemented in the multiphase flow and reactive transport simulator TOUGHREACT. DNF is simulated in two representative soils varying the application cycle, hydraulic loading rate, wastewater quality, water table depth, and subsurface heterogeneity. Simulations using the conventional specified flux boundary condition under-predict DNF by as much as 450% in sand and 230% in loamy sand compared to predictions from the coupled overland flow-vadose zone model, indicating that simulating coupled flow is critical for predicting DNF in cases where hydraulic loading rates are not sufficient to spread the wastewater over the whole basin. Smaller ratios of wetting to drying time and larger hydraulic loading rates result in greater water saturations, more anoxic conditions, and faster water transport in the vadose zone, leading to greater DNF. These results in combination with those from different water table depths explain why reported DNF varied with soil type and water table depth in previous field investigations. Across all simulations, cumulative percent DNF varies between 2 and 49%, indicating that NO3 removal in RIBS may vary widely depending on operational procedures and subsurface conditions. These modeling results improve understanding of DNF in RIBS and suggest operational procedures that may improve NO3 removal.
