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In recent decades, Crustacean cell line immortalization has drew a lot of attention, both academic and applied. We were trying to advance the state of art in this study by raising the proliferation rate of primary cells isolated from embryos of the massive freshwater prawn Macrobrachium rosenbergii by using a lentivirus that expresses the Ras oncogene. Ras is already expressed in silico testing of M. rosenbergii transcriptomic libraries for Ras expression, according to Complementarily, Ras is also expressed at very early stages of embryo formation. We transduced primary M. rosenbergii embryonic cells with a RasV12-expressing lentivirus by using the white spot syndrome virus promoter in the current study. In a subpopulation of cells with high granularity, the lenti-MrRas transduction rate was 23% in the total primary cell population and more than 80% in a subpopulation of cells with high granularity.
Source link: https://doi.org/10.3389/fmars.2022.1100971
When adding to the culturemedia, this research sought to determine the toxicity of a recreational dose of 60 bcg/mL of methamphetamine in vitro. Conclusions: Our results can be the first study to investigate the in vitro effects of methamphetamine on WJSCs at cellular level, resulting in a decrease in cell proliferation and viability when cells were exposed and treated with methamphetamine.
Source link: https://doi.org/10.30476/ijns.2022.97432.1210
Several cell manipulation experiments show that riboflavin, u00b7OH, and O2u2022-u2022-repeatible light exposure during regular cell manipulation is found here. These HEPES-tiary amine groups react with HEPA, resulting in 106. 6 bcM of H2O2. As the new generation biocompatible organic pH buffers, we have further developed and tested zwitterionic betaines, which are able to completely avoid the adverse effects that were found on HEPES and derivate Good's buffers. These findings may also open a new avenue for zwitterionic betaine-based betaine based materials for biomedical applications.
Source link: https://doi.org/10.1016/j.bioactmat.2022.12.028
Methods In an in vitro study, female menopausal rats were intragastrically administered quercetin at three doses for 90 days, and the estrous cycles were determined by vaginal smearing. At 5 bcM, 20 bcM, or 50 bbcM, rat primary ovarian granulosa cells were cultured and treated with H2O2 alone or H2O2+ quercetin in an in vitro study. By Western blot, the expression of the oxidative stress-related genes SOD-1, catalase, and glutathione synthetase were found in the ovaries and ovarian granulosa cells. In addition, the in vitro results showed that quercetin had remarkably reduced cell viability by H2u041e2-induced oxidative stress and raised the H2O2-induced decrease in the expression of oxidative stress-related proteins in vitro. Conclusions Both in vivo and in vitro, the results of this research indicated that quercetin improved the antioxidant capacity of the ovary by upregulating the expression of certain oxidative stress-related genes.
Source link: https://doi.org/10.1186/s13048-018-0421-0
Highly porous NFC hydrogel can be manufactured by freeze-drying NFC hydrogel. We showed that freeze-dried NFC hydrogel can be used for one-step 3D cell spheroid cell culturing of primary mesenchymal stem/stromal cells, prostate cancer cells, and hepatocellular carcinoma cells following the successful freeze-drying and reconstitution. No difference was found in the viability or morphology of the 3D cell spheroids cultured in the freeze-dried and reconstituted NFC hydrogel and fresh NFC hydrogel, respectively. These results show a step toward automatizing 3D cell culture and analysis, as well as an application to culture 3D cell spheroids more readily with the NFC hydrogel.
Source link: https://doi.org/10.3390/polym14245530
The blood-u2013brain barrier, which blocks the movement of substances into the brain, is created by brain microvascular endothelial cells. In vitro BBB models based on human-induced pluripotent stem cell-derived brain microvascular endothelial-like cells have been developed. This research was designed to produce iBMELCs from human iPS cell-derived endothelial progenitor cells. In the E-iBMELCs developed in this study, the endothelial cell markers' expression was higher relative to conventional methods. The E-iBMELCs developed in this research will greatly contribute to neurodegenerative diseases' discovery and may help with neurodegenerative diseases linked to BBB disruption.
Source link: https://doi.org/10.3390/pharmaceutics14122697
A complete PDMS micro-droplet chip for 3D cell culture was made by using SLA light-curing 3D printing techniques. The use of 3D printing techniques to produce micro-droplet chips can save manufacturing and time costs relative to the traditional preparation process of micro-droplet chips. The micro-droplet chip's different ratios of PDMS substrate and cover sheet, as well as the glue for making the convex mold can all increase the bonding strength and durability of the micro-droplet chip. The effectiveness of the complete PDMS micro-droplet chip in biological culture was tested by using a drug like chondrocyte suspension, and micro-droplet monitoring was achieved by controlling the flow rate of the dispersed phase and continuous phase. Experimental results show that the manufactured chip can satisfy experiments' demands, and it can be found that the micro-droplets of sodium alginate and the calcium chloride solution are cross-linked into three-dimensional microspheres.
Source link: https://doi.org/10.3390/mi13122050
The common two-dimensional cell culture techniques have a long tradition of imitating in vivo cell growth. Different mechanical stimuli that closely mimic the real in vivo microenvironment can be found in dynamic culture systems. The latest advancements in 3D dynamic cell culture techniques have been introduced in this series, with their advantages and disadvantages being addressed in comparison to traditional 2D cell culture in static conditions.
Source link: https://doi.org/10.3390/gels8120829
Boden membrane extracts obtained from Engelbreth sarcomas such as Matrigel u00ae remain the gold standard extracellular matrix for three-dimensional cell culture in cancer research. For 14 and 21 days of cell culture, respectively, MDA-MB-231 and MCF-7 breast cancer cell lines with various stiffnesses supported the growth and proliferation of MDA-MB-231 and MCF-7 breast cancer cell lines. Our results show that gelatin-based biomaterials provide an inexpensive and tunable 3D cell culture platform that can overcome traditional BMEs' limitations.
Source link: https://doi.org/10.3390/gels8120821
Summary: Primaris culture and long-term preservation of primary retinal pigment epithelium are a common model system for the analysis of ocular disease pathologies such as age-related macular degeneration. We explore the steps for the isolation and long-term culture of primary porcine RPE here. With immunofluorescence and histochemistry RPE cells grown on transwell membranes, we also discuss methods for cryoprotecting, cryosection, and interrogating.
Source link: https://doi.org/10.1016/j.xpro.2022.101758
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