Plant cell and organ cultures of Helianthella quinquenervis, a medicinal plant whose roots are used by the Tarahumara Indians of Chihuahua, Mexico,to relieve several ailments, were established to identify and quantify some chromenes with biological activity, such as encecalin, and to evaluate their potential for biotechnological production.
Gas chromatography-mass spectrometry (GC-MS) analysis corroborated the presence of quantifiable amounts of encecalin in H. quinquenervis cell cultures (callus and cell suspensions).
In addition, hairy roots were obtained through three transformation protocols (prick, 45-s sonication and co-culture), using wild type Agrobacterium rhizogenes A4.
After three months, cocultivation achieved the highest percentage of transformation (66%), and a comparable production (FW) of encecalin (110 μg/g) than the sonication assay (120 μg/g), both giving far higher yields than the prick assay (19 μg/g).
Stable integration of rolC and aux1 genes in the transformed roots was confirmed by polymerase chain reaction (PCR).
Hairy roots from cocultivation (six months-old) accumulated as much as 1086 μg/g (FW) of encecalin, over three times higher than the cell suspension cultures. The production of encecalin varied with growth kinetics, being higher at the stationary phase.
This is the first report of encecalin production in hairy roots of H. quinquenervis,demonstrating the potential for a future biotechnological production of chromenes.
Description: Delivers up to 150 Watts of ultrasonic power to the Titanium Tip. The Timer and Duty Cycle function increase preciosion in sample processing processing.
Description: Delivers up to 300 Watts of ultrasonic power to the Titanium Tip and includes an intergrated Sound Abating Chmaber to reduce cavitational sound emitted during processing. The Timer and Duty Cycle function increase preciosion in sample.
Description: Delivers up to 300 Watts of ultrasonic power to the Titanium Tip with preciosion control from a microprocessor and a graphical user interface displayed on a large (145 mm) LCD display. The integrated Sound Abating Chamber reduces cavitational sound emitted during processing.
Description: Designed to Perform multi-plate Assays on round 90/100mm Petri Dishes. The integrated LED illumination system provides transmitted light for brightfield and darkfield illumination of transparent media.
Description: A robotic liquid handling system designed to dispense Peni Cylinders and fill Peni Cylinders with the corresponding antibiotic liquid sample.
Custom development of ELISAs for other species or antibody isotypes not listed in the catalog. Custom testing of samples for IgG/IgM/IgA or total (IgG+IgM+IgA)
Intrinsic Type 1 Interferon (IFN1) Profile of Uncultured Human Bone Marrow CD45 low CD271 + Multipotential Stromal Cells (BM-MSCs): The Impact of Donor Age, Culture Expansion and IFNα and IFNβ Stimulation
Skeletal aging is associated with reduced proliferative potential of bone marrow (BM) multipotential stromal cells (MSCs). Recent data suggest the involvement of type 1 interferon (IFN1) signalling in hematopoietic stem cell (HSC) senescence.
Considering that BM-HSCs and BM-MSCs share the same BM niche, we investigated IFN1 expression profile in human BM-MSCs in relation to donor age, culture-expansion and IFN1 (α and β) stimulation.
Fluorescence-activated cell sorting was used to purify uncultured BM-MSCs from younger (19-41, n = 6) and older (59-89, n = 6) donors based on the CD45lowCD271+ phenotype, and hematopoietic-lineage cells (BM-HLCs, CD45+CD271–) were used as controls.
Gene expression was analysed using integrated circuits arrays in sorted fractions as well as cultured/stimulated BM-MSCs and Y201/Y202 immortalised cell lines.
IFN1 stimulation led to BM-MSC growth arrest and upregulation of many IFN1-stimulated genes (ISGs), with IFNβ demonstrating stronger effects. Uncultured MSCs were characterised by a moderate-level ISG expression similar to Y201 cells.
Age-related changes in ISG expression were negligible in BM-MSCs compared to BM-HLCs, and intracellular reactive oxygen species (ROS) levels in BM-MSCs did not significantly correlate with donor age. Antiaging genes Klotho and SIRT6 correlated with more ISGs in BM-MSCs than in BM-HLCs.
In patients with osteoarthritis (OA), BM-MSCs expressed considerably lower levels of several ISGs, indicating that their IFN1 signature is affected in a pathological condition. In summary, BM-MSCs possess homeostatic IFN1 gene expression signature in health, which is sensitive to in vitro culture and external IFN1 stimulation. IFN signalling may facilitate in vivo BM-MSC responses to DNA damage and combating senescence and aberrant immune activation.
Keywords: aging; bone marrow; mesenchymal stromal cells; senescence; type 1 interferon.
Ozone disinfection kinetics of poliovirus 1 determined by cellculture assay, RT-qPCR, and Ethidium Monoazide qPCR Reduction in a Continuous Quench-Flow Reactor
Aims: A continuous quench-flow (CQF) reactor was developed to collect samples at the reaction times of less than one second. The reactor is applied to determine ozone disinfection kinetics of poliovirus and to study whether EMA-qPCR can assess the viral infectivity after ozone disinfection.
Methods: Ozone disinfection of poliovirus was conducted in the developed CQF, and the disinfection kinetics were tested in the range of 0.7 – 5.0 seconds at ozone concentrationof 0.08 and 0.25 mg/L. Inactivation, damage on viral genome and damage on capsid integrity were determined by plaque assay, quantitative reverse transcription polymerase chain reaction (RT-qPCR), and ethidium monoazide treatment coupled with RT-qPCR (EMA-qPCR), respectively.
Postsynthetic Functionalization of DNA-Nanocomposites with Proteins Yields Bioinstructive Matrices for CellCulture Applications
We report on the directed postsynthetic functionalization of soft DNA nanocomposite materials with proteins. Using the example of the functionalization of silica nanoparticle-modified DNA polymer materials with agonists or antagonists of the epidermal growth factor receptor EGFR cell membrane receptor, we demonstrate that hierarchically structured interfaces to living cells can be established.
Due to the modular design principle, even complex DNA nanostructures can also be integrated into the materials, thereby enabling the high-precision arrangement of ligands on the lower nanometer length scale. We believe that such complex biohybrid material systems can be used for new applications in biotechnology.
Keywords: DNA nanocomposites; DNA origami nanostructure; epidermal growth factor receptor; phosphorylation; protein-DNA conjugates.
Human Muscle PrimaCell1: Normal Skeletal Muscle Cells Growth Medium
Description: Skeletal muscle cells are one of the largest cells in the body. They are multinucleate formed by the fusion of myoblasts. Skeletal muscle regeneration is a complex process in which many factors are involved. When skeletal muscle suffers an injury, quiescent resident myoblasts called satellite cells are activated to proliferate, migrate, and finally differentiate. Various cellular signaling pathways, such as phosphatidylinositol 3-kinase, calcineurin, Janus kinase 2/signal transducer and activator of transcription 3 (STAT3), and mitogen-activated protein kinase (MAPK) have been suggested to play an important role in skeletal muscle growth. RI-stimulated glucose transport in cultured human skeletal muscle is mediated by GLUT4 and heparan sulfate proteoglycan is involved in skeletal muscle differentiation. The fusion of mononucleated cells to form multinucleated myotubes is a central event in skeletal muscle development. Controlling the onset and progression of this process is a complex set of interactions between myoblasts and their environment. Skeletal muscle cell culture is a useful model for studying this process of cell differentiation.HSkMC from Gentaur Research Laboratories are isolated from human muscle of the pectoral girdle. HSkMC are cryopreserved at primary culture and delivered frozen. Each vial contains 5×10^5 cells in 1 ml volume. HSkMC are characterized by immunofluorescent method with antibodies to myosin, actin and actinin. HSkMC are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast and fungi. HSkMC are guaranteed to further expand for 15 population doublings at the conditions provided by Gentaur Research Laboratories.
Description: Human Skeletal Muscle Cells (HSkMC) are isolated from the limbal skeletal muscle. They are cryopreserved at second passage and can be cultured and propagated for at least 15 population doublings.
Tissue cDNA, First Strand, Human Adult Normal, Skeletal Muscle, BioGenomics
