November is National Alzheimer's Disease Awareness Month and research into a cure for the disease is vigorous. In a recently published paper "CRISPR/Cas9-Correctable mutation-related molecular andphysiological phenotypes in iPSC-derived Alzheimer’s PSEN2N141I neurons" researchers used the Celigo image cytometer upstream of CRISPR/Cas9 to visualize and asses cell death related to the sensitivity of iPSC-derived PSEN2N1411 neurons in the presence of Aβ42 oligomer toxicity.
The Novo Nordisk Center for Biosustainability (Denmark) set out to improve the efficiency of Chinese hamster ovary (CHO)-cell based production of non-monoclonal antibody, therapeutic glycoproteins designed to serve as biopharmaceuticals. To optimize the growth and production capacities of these CHO cells, the scientists looked at: lipid-based transfection, cell cultivation, cell counting, and antibody-independent product titer. Different growth and transfection parameters were investigated to see which yielded the highest growth profiles and production capacities. The Celigo was used in combination with Hoechst and propidium iodide to count the cells in 96-well format. The system developed here miniaturized the process and allowed [...]
The Notch-disrupting and cancer stem cell-inhibiting effects of the drug quinomycin A were investigated at the University of Kansas Medical Center. Using human pancreatic cancer cells PanC-1, MiaPaCa-2, and BxPC-3 and the Celigo to determine the number and size of pancreatospheres, researchers evaluated the drug’s ability to block cancer stem cell growth via inhibition of the Notch signaling pathway. After administration of the drug, proliferation and colony formation were blocked in cancer cell lines but not in normal pancreatic epithelial cells. Furthermore, cancer stem cell markers were reduced as was pancreatosphere formation. This work identifies quinomycin A as an efficacious [...]
The Danish Cancer Society Research Center recently published a study furthering their analysis of homologous recombination DNA repair machinery. The group previously reported on a growth factor, PSIP1, that enables DNA end resection. With GFP-transfected U2OS cells, the group investigated a structurally similar protein, hepatoma-derived growth factor-related protein 2 (HDGFRP2). The Celigo analyzed cell number and viability via fluorescent markers. The group reports that HDGRFP2 may help to repair silent genes that have been impaired or active genes inhibited by DNA damage. Read the full publication here.
At the Canary Center at Stanford for Early Cancer Detection, investigators studied how AshwaMAX (a steroidal lactone from a winter cherry plant, Withania somnifera, extract) might work as an oral treatment for those with the highly aggressive cancer glioblastoma multiforme (GBM). A heterogeneous disease, non-specific therapies for GBM have proven largely ineffective. Two patient-derived GBM lines (GBM2, GBM39) and one GBM cell line were cultured to create neurospheres that were then exposed to various concentrations of AshwaMAX. Celigo measured cell proliferation and cell death via Trypan Blue staining. AshwaMAX inhibited the neurospheres at nanomolar concentrations. After additional work in vivo, [...]
Here's a great example of how the Celigo image cytometer is able to perform common experiments while saving time and money! Ignyta, Inc. was looking for a new way to perform reagent-free proliferation analyses with suspension cells. This new method had to produce results which correlated well to their current method, Cell Titer-Glo®. Nexcelom and Ignyta partnered to perform a head-to-head cell proliferation comparison between Celigo® and Cell Titer-Glo. Using four suspension cell types (Ba/F3 parental cell line, Ba/F3 expressing an oncogenic gene, oncogenic gene mutant A and B), Ignyta plated all cells at a concentration of 5,000 cells/well in [...]
Introduction Chinese hamster ovary (CHO) cells are the cell type of choice for biopharmaceuticals due to their propensity to correctly fold and post-translationally modify human proteins . Here, researchers use an optimized bioinformatics tool (“CRISPy”) to generate chimeric RNA called “single guide” RNA (sgRNA) in order to disrupt FUT8, a gene that is utilized in N-glycosylation. This novel web-based tool ensures efficient, fast, and low-cost genetic manipulation of CHO cells for future biopharmaceutical applications. Materials and Methods Cell culture and transfection of CHO cells CHO cells were cultured and transfected by Nucleofector Kit V with 1 μg of Cas9 plasmid [...]
Investigation of IAPP Role in Increasing ROS Production and Apoptosis in p53-deficient Tumor Cells using Celigo Imaging Cytometer
The entire family of tumor protein p53 (TP53) enhances functions such as apoptosis and autophagy in normal cellular functioning. TP53 is a tumor repressor gene that is often inactivated in human cancers. Reactivating p53 has proven difficult to achieve therapeutically, however. Researchers at MD Anderson Cancer Center are investigating other members of the p53 pathway in order to elucidate new therapeutic options to suppress p53-deficient tumor growth. ΔN isoforms of two members of the p53 family, p63 and p73, are usually overexpressed in cancers and these isoforms (which lack the acidic transactivation domain) act on p53 in a dominant-negative fashion, [...]