Cell Signaling and Neuroscience
The ADSCs growing on the microcarriers and those inmonolayer culture were harvested, digested and smeared. They werethen subjected to safranin-O and toluidine blue staining to observethe levels of glycosaminoglycan (GAG) synthesis and secretion fromthe extracellular matrix. The type II collagen monoclonal antibodywas incubated with the cells with DAB (Beijing Hepten and ProteinBiomedical Institute, Beijing, China) as the substrate, and theimmunoperoxidase method was used for development. Brown stainingindicated positive staining, and thus the occurrence of type IIcollagen secretion and synthesis.
Vitamin B3 (Niacin) - Scientific Review on Usage, …
N2 - Background: Carcinoid tumors are associated with the carcinoid syndrome, a set of symptoms resulting from the peptide and amine products, including serotonin, secreted from the cancer cells. The purpose of this study was to investigate the relationship between the phosphatidylinositol-3-kinaselprotein kinase B (PI3K/Akt) inhibitor PTEN (phosphatase and tensin homolog deleted on chromosome ten) and serotonin synthesis and secretion in the carcinoid cancer cell line BON. Materials and Methods: PTEN was inhibited by pharmacological and molecular approaches, and the resultant secretion of serotonin and expression of tryptophan hydroxylase 1 (TPH1), the rate-limiting enzyme in serotonin synthesis, was assessed. Results: Inhibition of PTEN in vitro, with concomitant increased Akt signaling, resulted in decreased secretion of serotonin, as well as decreased serotonin synthesis, as confirmed by reduced expression of TPH1. Inhibition of PTEN in BON cells in an animal model resulted in decreased serum serotonin. Conclusion: By inhibiting signaling through Akt, PTEN indirectly promotes serotonin synthesis and secretion.
Human ADSCs were cultured for 1, 3, 5 and 7 days,and under the inverted microscope, the morphology of the ADSCs andthe proliferation on the surface of the microcarriers wereassessed. The chondrocytes on the surface of the microcarrier weredigested, separated and counted with a blood cell counting plate(Yancheng Glass Instrument Company, Yancheng, China), then a cellgrowth curve was constructed. Cell microcarrier samples were takenon day 3 to be observed by scanning electron microscopy (HitachiS-4500; Hitachi, Tokyo, Japan), and the cell morphology and matrixsecretion was observed. The static monolayer culture of ADSCs inthe control group was observed to assess the ADSC morphology andproliferation at days 1, 3, 5 and 7. They were then collected andthe survival rate was calculated using a trypan blue dye exclusiontest. Finally, a cell growth curve was constructed based on wholecell counts.
Dissociation of IL-1 beta synthesis and secretion in …
The Cytodex 3 microcarriers (50 mg) and ADSCs (finalconcentration, 1×10/ml) were added to a 10-ml rotarycell culture system (RCCS; Synthecon, Inc., Houston, TX, USA)culture container, then the cartilage inducing medium was added tofill the container. The container with cell-microcarriers complexwas connected to the rotating base. The device was put in a 5%CO incubator and was cultured at 37°C. In order tofully mix cells, the container was rotated at a force of 5 rpm for5 min and paused for 5 min alternately, which was repeated forseveral cycles, it was then continuously rotated and graduallyadjusted to 10–12 rpm. Once the culture medium exhibited a yellowcolor and a PH value /ml ADSCs intofive wells of a 6-well plate, 2 ml in each well. During theculture, the medium was changed every 2–3 days to maintain asufficient quantity of cells.