Primary Cell e Newsletter Issue 18 (jun 2016) Cell Applications

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Traffic Jam at the Blood–Brain Barrier Promotes Greater Accumulation of Alzheimer’s Disease Amyloid-β Proteins in the Cerebral Vasculature Molecular Pharmaceutics, 10:1557–1565 Highlight Study Primary Cell Biology News Issue 18 Jun 2016 Primary Cells Media & Reagents Cell Biology Starting Material Matters - Cell signaling pathway strength may vary among donors, so researchers normally must set aside time to screen cells for sensitivity to specific stimuli. Investigators can save valuable resources 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 signaling pathways. Vascular Endothelial Growth Factor Receptor (VEGFR-2)-mediated signaling plays a critical role in endothelial cell proliferation, differentiation, movement and survival, as well as vessel formation. In this pathway, VEGF-induced receptor dimerization 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-Based Assays for Drug Toxicity and Genotoxicity Testing Using automated high-res microscopy & analytical software to visualize and quantify cell responses, HCS determines how compounds affect cell morphology in a rapid, quantitative, high-throughput manner 2. An Intro To HCS: Imaging Technology, Assay Development and Data Analysis in Biology & Drug Discovery A collaborative and interdisciplinary approach covering HCS concepts, principles, training and tutorials for pharmaceutical industry and academia 3. Assay Development Guidelines for Image-Based High Content Screening, Analysis and Imaging Immunofluorescence, confocal and wide-field microscopy with automation, image analysis and quantitative information to detect multiple changes in single cells. Used for target ID, compound profiling & development. Primary cell assays exploit natural genetics, reactions, assembly & physiology. 4. Image-Based High-Content Screening in Drug Discovery Automated, microscope-based HCS looks at many features simultaneously in complex biology systems. HCS is used in the pharmaceutical drug discovery pipeline to validate drug targets & compounds, predict toxicity, and suggest pathways or molecular targets. High Content Screening Browse Past Issues 2 1 3 4 Pre-Screened for Angiogenesis & VEGF Signaling The Figure at right shows activation of VEGFR-2 signaling in endothelial cells from human aorta (HAOEC), umbilical vein (HUVEC) and microvessels (HMVEC) by 100 ng/ml VEGF for 0, 5 or 15 minutes. VEGFR-2 autophosphorylation is probed with phospho-VEGFR-2 specific antibody, while Akt activation is detected 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 & HMVEC express key endothelial markers Tie2, eNOS, Axl and Etk/Bmx. Amyloid-β40 (Aβ40) deposition in the brain vasculature results in cerebral amyloid angiopathy (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 findings uncover pathophysiological events that promote CAA and suggest clearing vascular endothelium of Aβ proteins may halt or even reverse CAA. 1. MicroRNA suppresses SMC differentiation involved in heart disease Useful Links and Downloads Dr. Anna Wakeland joins CAI as Director of Technical Sales Now Trending: 3. Cellular Signaling 2. IEEE Transactions Nanotech 1. PLoS ONE 3. Downstream signaling pathways of common growth factor described. Excessive vascular smooth muscle cell proliferation contributes to pulmonary blockage and hypertension. MicroRNA suppresses gene signals involved in differentiation of human pulmonary artery SMCs. 2. Real-time biological cell sensing system based on carbon nanofibers Non-invasive, label-free measurement of cell behavior & response: Changes in cell morphology & integrity provide clues for toxicity, disease diagnosis, environmental monitoring & drug effectiveness. Hepatocyte Growth Factor isoform induces cell motility & survival: HGF Isoform NK2 was studied in primary Human Bronchial Epithelial Cells HPASMC HBEpC BAOSMC Primary Cells: Human PBMC Femoral Artery Cells Neural 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 direct sales, 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, she mastered comprehensive human stem cell culture techniques, developed protocols for differentiation of human pluripotent stem cells and analyzed trophoblast lineage specification. Prior roles included a research internship at Sandia National Laboratories. Dr. Wakeland earned her PhD in Biomedical Sciences from UCSD and completed a Micro-MBA Certificate Program at the Rady School of Management. She also participated in a Biotechnology Life Science Business Immersion Program through the BIOCOM Institute. “We are excited about the scientific and professional skills Dr. Wakeland brings to the commercial life science setting," says Daniel Schroen, Vice President of Sales and Marketing at CAI. "With her business acumen and deep knowledge of primary cell biology, Anna will be able to help lab researchers select optimal products and services to successfully complete even the most intricate, large scale projects.” CD34+ Progenitors Cytofect SMC Trasnfection Kit and Anna Wakeland, PhD and Bronchial / Tracheal Epitheal Cell Medium Blood-Brain Barrier Digestive & Liver Disease, June 2014 PLoS ONE, May 2016 cellapplications.com Limited Time - 40% OFF Bovine Brain Microvascular Endothelial Cells (BBMVEC) What are integration- free Human iPSC? F A Q BBMVEC Coupon Code* *Enter Coupon Code at check out online, or mention code when ordering by phone, e-mail or fax. A Classic System to Study the Blood-Brain Barrier (BBB) Integration-free Human Induced Pluripotent Stem Cells (HiPSC) rely on strategies to enforce the expression of reprogramming factors in the host cells that do not imply the insertion of vector DNA in the host genome. Specifically, the Sendai virus is an RNA virus that never goes through a DNA phase, thus there is no risk of random integration of viral genetic material in the host genome. The pioneering work describing the generation of HiPSCs utilized retroviruses to transfer the reprogramming factors to donor skin fibroblasts. An effective strategy, the use of retrovirus implies that the vriuses’ genetic material is integrated in host genome, along with the transgenes. This somewhat uncontrolled, random integration could potentially cause genotoxic stress or affect cell behavior genes. Thus, integration-free HiPSC can help ensure an observed phenotype is the result of a specific genetic mutation or drug. The Blood-Brain Barrier in 3D Media Contact Daniel Schroen, PhD VP, Sales & Marketing (858) 453-0848 [email protected] Rat Brain Microvascular Endothelial Cells (RBMVEC) Expires June 30, 2016

Transcript of Primary Cell e Newsletter Issue 18 (jun 2016) Cell Applications

Page 1: 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

21

3 4

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

Limited Time - 40% OFF

Bovine Brain MicrovascularEndothelial Cells (BBMVEC)

What are integration-free Human iPSC?

F A Q

BBMVEC

Coupon Code*

*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

[email protected]

Rat Brain Microvascular Endothelial Cells (RBMVEC)

Expires June 30, 2016