Primary Cell e Newsletter Issue 18 (jun 2016) Cell Applications
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Transcript of Primary Cell e Newsletter Issue 18 (jun 2016) Cell Applications
Traffic Jam at the Blood–Brain Barrier Promotes Greater Accumulation ofAlzheimer’s Disease Amyloid-β Proteins in the Cerebral Vasculature
Molecular Pharmaceutics, 10:1557–1565Highlight Study
Primary Cell Biology NewsIssue 18 Jun 2016
Primary Cells Media & Reagents Cell Biology
Starting Material Matters - Cell signaling pathway strength may vary among donors, so researchers normallymust set aside time to screen cells for sensitivity to specific stimuli. Investigators can save valuableresources and improve the validity and reproducibility of results through the use of pre-screened cells.
Bovine Brain Microvascular Cells (BBMVEC) were used in this study
Endothelial cells can be pre-screened to demonstrate stimulation-dependent angiogenesis and key endothelial cell signalingpathways. Vascular Endothelial Growth Factor Receptor(VEGFR-2)-mediated signaling plays a critical role in endothelialcell proliferation, differentiation, movement and survival, as well asvessel formation. In this pathway, VEGF-induced receptordimerization triggers activation of VEGFR-2 tyrosine kinase,leading to activation of downstream signaling molecules.
Pre-screening helps normalize cell signaling responsiveness to target stimuli
BBMVEC
Human Endothelial Cells:
1. High-Content Screening and Analysis in Cell-BasedAssays for Drug Toxicity and Genotoxicity TestingUsing automated high-res microscopy & analytical software to visualize andquantify cell responses, HCS determines how compounds affect cellmorphology in a rapid, quantitative, high-throughput manner
2. An Intro To HCS: Imaging Technology, Assay Developmentand Data Analysis in Biology & Drug DiscoveryA collaborative and interdisciplinary approach covering HCS concepts,principles, training and tutorials for pharmaceutical industry and academia
3. Assay Development Guidelines for Image-Based HighContent Screening, Analysis and Imaging
Immunofluorescence, confocal and wide-field microscopy with automation,image analysis and quantitative information to detect multiple changes in singlecells. Used for target ID, compound profiling & development. Primary cellassays exploit natural genetics, reactions, assembly & physiology.
4. Image-Based High-Content Screening in Drug Discovery
Automated, microscope-based HCS looks at many featuressimultaneously in complex biology systems. HCS is used in thepharmaceutical drug discovery pipeline to validate drug targets &compounds, predict toxicity, and suggest pathways or molecular targets.
High Content Screening
Browse
Past Issues
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Pre-Screened for Angiogenesis & VEGF Signaling
The Figure at right shows activation of VEGFR-2 signaling inendothelial cells from human aorta (HAOEC), umbilical vein(HUVEC) and microvessels (HMVEC) by 100 ng/ml VEGF for 0, 5or 15 minutes. VEGFR-2 autophosphorylation is probed withphospho-VEGFR-2 specific antibody, while Akt activation isdetected by a phospho-Akt. A phospho-p44/42 MAP kinase(T202/Y204) antibody is used to demonstrate kinase activation.
The bottom panel illustrates that pre-screened HUVEC & HMVECexpress key endothelial markers Tie2, eNOS, Axl and Etk/Bmx.
Amyloid-β40 (Aβ40) deposition in the brain vasculature results in cerebral amyloidangiopathy (CAA), which occurs in Alzheimer’s disease. Dutch Mutation of Aβ40(E22Q) in carriers promotes accumulation in the endothelial cells of the blood–brain-barrier (BBB). The resulting permeation barrier prevents Amyloid clearance,promotes deposits in microvessels, and leads to severe CAA. These findingsuncover pathophysiological events that promote CAA and suggest clearing vascularendothelium of Aβ proteins may halt or even reverse CAA.
1. MicroRNAsuppresses SMC
differentiation involvedin heart disease
Useful Links and Downloads
Dr. Anna Wakeland joins CAI as Director of Technical Sales
Now Trending:
3. Cellular Signaling2. IEEE Transactions Nanotech1. PLoS ONE
3. Downstreamsignaling pathways of
common growthfactor described.
Excessive vascular smooth muscle cell proliferationcontributes to pulmonary blockage and hypertension.MicroRNA suppresses gene signals involved indifferentiation of human pulmonary artery SMCs.
2. Real-time biologicalcell sensing system
based on carbonnanofibers
Non-invasive, label-free measurement of cellbehavior & response: Changes in cell morphology &integrity provide clues for toxicity, disease diagnosis,environmental monitoring & drug effectiveness.
Hepatocyte Growth Factor isoform induces cellmotility & survival: HGF Isoform NK2 was studiedin primary Human Bronchial Epithelial Cells
HPASMC
HBEpC
BAOSMC
Primary Cells:
Human PBMCFemoral Artery CellsNeural Stem Cells
MicroRNA, Biological Cell Sensing, Growth Factor Signaling
May 23, 2016 - Cell Applications, Inc. is pleased to announce that Anna Wakeland,PhD, has joined the company as Director of Technical Sales. In this key role, Dr.Wakeland will leverage her strong research and presentation skills to grow directsales, providing solution-centric, consultative approaches to the customer experience. Most recently, Anna conducted cell-based research at the University of California,San Diego, including an interim role as Clinical Research Associate. At UCSD, shemastered comprehensive human stem cell culture techniques, developed protocolsfor differentiation of human pluripotent stem cells and analyzed trophoblast lineagespecification. Prior roles included a research internship at Sandia NationalLaboratories. Dr. Wakeland earned her PhD in Biomedical Sciences from UCSD andcompleted a Micro-MBA Certificate Program at the Rady School of Management. She also participated in a Biotechnology Life Science Business Immersion Programthrough the BIOCOM Institute. “We are excited about the scientific and professionalskills Dr. Wakeland brings to the commercial life science setting," says DanielSchroen, Vice President of Sales and Marketing at CAI. "With her business acumenand deep knowledge of primary cell biology, Anna will be able to help lab researchersselect optimal products and services to successfully complete even the most intricate,large scale projects.”
CD34+ Progenitors
Cytofect SMCTrasnfection Kit
and
Anna Wakeland, PhD
andBronchial /
Tracheal EpithealCell Medium
Blood-Brain Barrier
Digestive & Liver Disease, June 2014
PLoS ONE, May 2016
cellapplications.com
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Bovine Brain MicrovascularEndothelial Cells (BBMVEC)
What are integration-free Human iPSC?
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*Enter Coupon Code at check out online, or mentioncode when ordering by phone, e-mail or fax.
A Classic System to Study the Blood-Brain Barrier (BBB)
Integration-free Human Induced Pluripotent StemCells (HiPSC) rely on strategies to enforce theexpression of reprogramming factors in the hostcells that do not imply the insertion of vector DNAin the host genome. Specifically, the Sendai virusis an RNA virus that never goes through a DNAphase, thus there is no risk of random integrationof viral genetic material in the host genome.
The pioneering work describing the generation ofHiPSCs utilized retroviruses to transfer thereprogramming factors to donor skin fibroblasts.An effective strategy, the use of retrovirus impliesthat the vriuses’ genetic material is integrated inhost genome, along with the transgenes. Thissomewhat uncontrolled, random integration couldpotentially cause genotoxic stress or affect cellbehavior genes. Thus, integration-free HiPSC canhelp ensure an observed phenotype is the result ofa specific genetic mutation or drug.
The Blood-Brain Barrier in 3D
Media ContactDaniel Schroen, PhD
VP, Sales & Marketing(858) 453-0848
Rat Brain Microvascular Endothelial Cells (RBMVEC)
Expires June 30, 2016