What process is used to obtain cells from bone marrow and normal peripheral blood?
What is the best cell counting and viability method for primary cells?
AllCells, your primary cells research partner, and Nexcelom Bioscience, your cell counting experts, have joined together in an exclusive collaboration to host a free webinar to help educate researchers and present data from their own experiences.
Selecting a model system is usually one of the first and most challenging steps in exploring a clinical hypothesis, validating a technology, or understanding a biological process. Although cell lines have served the needs of biomedical research for decades, increasingly, grant and publication reviewers as well as agencies regulating drug development, are requesting researchers to reproduce their results in more representative or “clinically relevant” models. This webinar will introduce human and animal-derived primary cells and explain how they currently serve diverse biomedical clinical and research applications as biologically-relevant, species-specific, and simple to use biological systems.
Cell Storage Best Practices: Techniques to Ensure Greater Research Resultsallcells
Featuring: Cryopreservation Specialist, Michael George
This webinar will introduce cell storage equipment and techniques in order to provide the most up-to-date trends in research today. There will be special emphasis on human primary cell storage, cryopreservation, and transportation.
Topics to be covered in the webinar:
• Background on life science storage methods
• Advantages/disadvantages of storage equipment
• What not to do (common mistakes/misconceptions)
• Interview with James Lee, Associate Director of Production, AllCells
• Q&A
This slide is all about the hematopoeitic stem cells its two types myeloid and lymphoid. The different types of myleoid and lymphoid cells are explained in details. All details about different White Blood Cells and their function. B cell, T cell and Natural Killer cell and their function.
Selecting a model system is usually one of the first and most challenging steps in exploring a clinical hypothesis, validating a technology, or understanding a biological process. Although cell lines have served the needs of biomedical research for decades, increasingly, grant and publication reviewers as well as agencies regulating drug development, are requesting researchers to reproduce their results in more representative or “clinically relevant” models. This webinar will introduce human and animal-derived primary cells and explain how they currently serve diverse biomedical clinical and research applications as biologically-relevant, species-specific, and simple to use biological systems.
Cell Storage Best Practices: Techniques to Ensure Greater Research Resultsallcells
Featuring: Cryopreservation Specialist, Michael George
This webinar will introduce cell storage equipment and techniques in order to provide the most up-to-date trends in research today. There will be special emphasis on human primary cell storage, cryopreservation, and transportation.
Topics to be covered in the webinar:
• Background on life science storage methods
• Advantages/disadvantages of storage equipment
• What not to do (common mistakes/misconceptions)
• Interview with James Lee, Associate Director of Production, AllCells
• Q&A
This slide is all about the hematopoeitic stem cells its two types myeloid and lymphoid. The different types of myleoid and lymphoid cells are explained in details. All details about different White Blood Cells and their function. B cell, T cell and Natural Killer cell and their function.
Get the bone marrow transplant treatment in Delhi by best doctor / surgeon in world class hospital. We ensure you get the best medical care and even stay in touch for more assistance. website :- http://www.transplantsurgeryindia.com/bone-marrow-transplants/
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
This presentation deals with stem cell therapy & new avenues in stem cell therapy. It also discusses latest advances such as treatment against baldness, multiple sclerosis, type 1 diabetes, spinal cord injury, demyelinating diseases, deafness, eye, Parkinson's disease. Also discusses about umbilical cord stem cells and finally clinical trials without patients (organs on chips).
What is Stem Cell ?
History of Stem Cells ?
Stages of Embryogenesis
Blastocyst Diagram
Three types of stem cells
Differentiation of ESC
Adult Stem Cells
Bone Marrow
Umbilical cord stem cells
Factors known to affect stem cells
Niche cells activates Stem cells
Regenerative Medicine : Indian Scenario
Get the bone marrow transplant treatment in Delhi by best doctor / surgeon in world class hospital. We ensure you get the best medical care and even stay in touch for more assistance. website :- http://www.transplantsurgeryindia.com/bone-marrow-transplants/
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
This presentation deals with stem cell therapy & new avenues in stem cell therapy. It also discusses latest advances such as treatment against baldness, multiple sclerosis, type 1 diabetes, spinal cord injury, demyelinating diseases, deafness, eye, Parkinson's disease. Also discusses about umbilical cord stem cells and finally clinical trials without patients (organs on chips).
What is Stem Cell ?
History of Stem Cells ?
Stages of Embryogenesis
Blastocyst Diagram
Three types of stem cells
Differentiation of ESC
Adult Stem Cells
Bone Marrow
Umbilical cord stem cells
Factors known to affect stem cells
Niche cells activates Stem cells
Regenerative Medicine : Indian Scenario
In the development of cancer immunotherapies, vaccines, and autoimmune disease treatments, researchers are increasingly aware of the importance of host human leukocyte antigens (HLA) in understanding the complexity of immune responses. HLA allelic variants have distinct binding affinities for peptide antigens during presentation, and thus regulate auto-antigen and pathogen-specific T cell activation. HLA polymorphisms have been correlated with disease risk, immune responses to infectious disease, as well as adverse responses to therapies. In this webinar, we will review the genetics and biology of HLA types and explain the technology underlying HLA identification. We will also provide some examples of how HLA-typing can be a valuable investment during early stage drug-development in human primary cells.
Purification of total RNA from peripheral blood mononuclear cells - Download ...QIAGEN
Peripheral blood is often used for in vitro studies of the human immune system or immune responses, such as inflammation. An important part of the human immune system is represented by the peripheral blood mononuclear cells (PBMC). PBMC are blood cells characterized by a round nucleus and consist mainly of lymphocytes (T cells, B cells, and NK cells), macrophages and dendritic cells. Here, we describe the analysis of lipopolysaccharide-induced transcriptional response of isolated PBMC from whole blood using the RNeasy® Mini Kit or RNeasy Micro Kit, RT2 First Strand Kit, RT2 SYBR® Green ROX™ qPCR Mastermix, and RT2 Profiler PCR Arrays.
Critical Steps for Real-Time PCR Analysis: Tips and Solutions to Achieve Effi...QIAGEN
In this slidedeck, we cover the following topics which are critical steps for efficient and precise gene expression studies using real-time PCR technology:
1) Effect of RNA integrity on real-time PCR results – tips to achieve a true RNA profiling suitable for real-time PCR studies
2) Improved methods for cDNA synthesis, optimized for real-time PCR
3) Real-time PCR analysis:
• Real-time PCR essentials and background information on different quantification strategies
• SYBR Green real-time PCR – factors influencing specificity
• Introduction to probe technology
• New, fast and efficient real-time PCR solutions
Better cell cultures lower your research costs!-A common Neuromics' theme is harnessing the power of cells. The raw cost of the cells are often the biggest consideration. We encourage our customers to focus on true costs. These include the # number of cells (how many times can they be passaged?), culture viability (how long do the cells live) and bioactivity (how closely do cultures mimic in vivo behavior?). I would like to present a publication and presentation confirms our competitiveadvantage when analyzing true costs.
Specialty pathology endpoints for all clients, including complex immunohistochemistry, in situ hybridization, image analysis, artificial intelligence, stereology, tissue cross-reactivity, and electron microscopy.
This is my short presentation in one of my university classes. It's obvious that the future of the stem cell biology is tightly engaged with organoids and they will absolutely change the way science is going to.
Kind regards
Shahin Ahmadian
Human Cell Systems Biology for Drug Discovery and Chemical Safety. Presentation at the 7th Brazilian Symposium on Medicinal Chemistry, November 12, 2014, Campos do Jordao-SP, Brazil. Ellen Berg.
LF Energy Webinar: Electrical Grid Modelling and Simulation Through PowSyBl -...DanBrown980551
Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
Then welcome to this PowSyBl workshop, hosted by Rte, the French Transmission System Operator (TSO)!
During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
- A fully editable and extendable library for grid component modelling;
- Visualization tools to display your network;
- Grid simulation tools, such as power flows, security analyses (with or without remedial actions) and sensitivity analyses;
The framework is mostly written in Java, with a Python binding so that Python developers can access PowSyBl functionalities as well.
What you will learn during the webinar:
- For beginners: discover PowSyBl's functionalities through a quick general presentation and the notebook, without needing any expert coding skills;
- For advanced developers: master the skills to efficiently apply PowSyBl functionalities to your real-world scenarios.
GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
Key Trends Shaping the Future of Infrastructure.pdfCheryl Hung
Keynote at DIGIT West Expo, Glasgow on 29 May 2024.
Cheryl Hung, ochery.com
Sr Director, Infrastructure Ecosystem, Arm.
The key trends across hardware, cloud and open-source; exploring how these areas are likely to mature and develop over the short and long-term, and then considering how organisations can position themselves to adapt and thrive.
Transcript: Selling digital books in 2024: Insights from industry leaders - T...BookNet Canada
The publishing industry has been selling digital audiobooks and ebooks for over a decade and has found its groove. What’s changed? What has stayed the same? Where do we go from here? Join a group of leading sales peers from across the industry for a conversation about the lessons learned since the popularization of digital books, best practices, digital book supply chain management, and more.
Link to video recording: https://bnctechforum.ca/sessions/selling-digital-books-in-2024-insights-from-industry-leaders/
Presented by BookNet Canada on May 28, 2024, with support from the Department of Canadian Heritage.
Connector Corner: Automate dynamic content and events by pushing a buttonDianaGray10
Here is something new! In our next Connector Corner webinar, we will demonstrate how you can use a single workflow to:
Create a campaign using Mailchimp with merge tags/fields
Send an interactive Slack channel message (using buttons)
Have the message received by managers and peers along with a test email for review
But there’s more:
In a second workflow supporting the same use case, you’ll see:
Your campaign sent to target colleagues for approval
If the “Approve” button is clicked, a Jira/Zendesk ticket is created for the marketing design team
But—if the “Reject” button is pushed, colleagues will be alerted via Slack message
Join us to learn more about this new, human-in-the-loop capability, brought to you by Integration Service connectors.
And...
Speakers:
Akshay Agnihotri, Product Manager
Charlie Greenberg, Host
"Impact of front-end architecture on development cost", Viktor TurskyiFwdays
I have heard many times that architecture is not important for the front-end. Also, many times I have seen how developers implement features on the front-end just following the standard rules for a framework and think that this is enough to successfully launch the project, and then the project fails. How to prevent this and what approach to choose? I have launched dozens of complex projects and during the talk we will analyze which approaches have worked for me and which have not.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
Let's dive deeper into the world of ODC! Ricardo Alves (OutSystems) will join us to tell all about the new Data Fabric. After that, Sezen de Bruijn (OutSystems) will get into the details on how to best design a sturdy architecture within ODC.
Essentials of Automations: Optimizing FME Workflows with ParametersSafe Software
Are you looking to streamline your workflows and boost your projects’ efficiency? Do you find yourself searching for ways to add flexibility and control over your FME workflows? If so, you’re in the right place.
Join us for an insightful dive into the world of FME parameters, a critical element in optimizing workflow efficiency. This webinar marks the beginning of our three-part “Essentials of Automation” series. This first webinar is designed to equip you with the knowledge and skills to utilize parameters effectively: enhancing the flexibility, maintainability, and user control of your FME projects.
Here’s what you’ll gain:
- Essentials of FME Parameters: Understand the pivotal role of parameters, including Reader/Writer, Transformer, User, and FME Flow categories. Discover how they are the key to unlocking automation and optimization within your workflows.
- Practical Applications in FME Form: Delve into key user parameter types including choice, connections, and file URLs. Allow users to control how a workflow runs, making your workflows more reusable. Learn to import values and deliver the best user experience for your workflows while enhancing accuracy.
- Optimization Strategies in FME Flow: Explore the creation and strategic deployment of parameters in FME Flow, including the use of deployment and geometry parameters, to maximize workflow efficiency.
- Pro Tips for Success: Gain insights on parameterizing connections and leveraging new features like Conditional Visibility for clarity and simplicity.
We’ll wrap up with a glimpse into future webinars, followed by a Q&A session to address your specific questions surrounding this topic.
Don’t miss this opportunity to elevate your FME expertise and drive your projects to new heights of efficiency.
Leading Change strategies and insights for effective change management pdf 1.pdf
Isolating Primary Cells 101
1. Isolating Primary Cells 101: Which viability
method is best for your research?
Ask the Experts Series #1
AllCells - Nexcelom Bioscience
Exclusive Webinar Event
2/13/2014
James Lee and Leo Chan, Ph.D.
2. Company Information
• Provider of image
cytometry technology
for cell concentration
and viability
• Leo Chan, Ph.D.
• Provider of primary
cells & bioservices for
pre-clinical research
• James Lee
3. Important Questions to Address
• What is the process
used for obtaining cells
from bone marrow?
• What is the process
used for obtaining cells
from peripheral blood?
• What is the best and
most accurate cell
counting and viability
method for primary
cells?
• How do we select the
appropriate method for
viability measurement?
4. Key Points to Take Away
• AllCells key takeaways: decrease variability, increase speed and
accuracy
• Nexcelom Bioscience key takeaways: the best method for
concentration and viability measurement using different
Cellometers, which is cell sample dependent
5. Company Information
ALLCELLS is a leading B2B provider of hematology-based cells & services
to the world’s life science research community for the past 16 years.
Products: >800 healthy & diseased human tissue and primary cell
products that enable drug discovery, stem cell research, and
advancement of therapies
Bioservices: Applications of hematology /
immunology expertise; cell-based assay
services and customized research studies
Core Competencies: Hematology, immunology, human tissue
procurement (healthy & diseased), large-scale cell isolations &
characterization, stem cell expansion & differentiation
Operations: Alameda, CA (HQ), IRB approved collection
programs, Tissue Bank and Biologics Licenses
6. Primary Cells - Uses
•
•
•
•
•
Drug Development – Pre-clinical Development
Predictive Toxicology
Stem Cell Research/Development
Regenerative Medicine
Medical Device Validation
7. Primary Human Cells by AllCells
• Normal Peripheral Blood -> MNC -> T Cells
(untouched)
• Bone Marrow -> MNC -> CD34+
(organ>isolation steps-> cell types)
(Cell viability, counting methods suitable for
each step)
10. Whole Bone Marrow
Aspirate Information
Whole Bone Marrow is drawn into a 10cc syringe containing
heparin (80U/ml of BM) from the posterior iliac crest,
25ml/site, from a maximum of four sites.
11. Whole Bone Marrow
• Whole Bone Marrow -> CD34+ and CD133+ isolation methods
• Small/large scale isolations
• Fluorescence staining
- Trypan blue method
- AOPI method
• Cryopreservaton Method
Post Thaw Data
Date
Tissue
Type/Unit
Lot Number
AO/PI Lot
Number
Cellometer
Count
Cellometer
Viability
Hemacytometer
Count
Trypan Blue
Viability
11-20-13 ABM-CD34+
0.5x10^6
BM4702
130904-01
0.8x10^6
97%
0.9x10^6
90%
11-20-13 ABM-CD34+
0.1x10^6
BM4699
130904-01
0.3x10^6
92%
0.3x10^6
96%
12. Normal Peripheral Blood
Leuko PakTM
Leuko Pak is PBMCs collected from donors by
apheresis, from Ancient Greek ἀφαίρεσις (aphairesis,
“a taking away”) is a medical technology in which the
blood of a donor or patient is passed through an
apparatus that separates out one particular
constituent and returns the remainder to the
circulation. It is thus an extracorporeal therapy. The
apparatus we us is COBE Spectra Apheresis System
manufactured by Caridian BCT. The separation of
individual blood component is done with a specialized
centrifuge. Spectra system may also be used to collect
platelets, plasma and granulocytes.
Approximately three donor blood volume is processed
and between 250-300ml of plasma and MNC is
collected/donor.
•Average cell yield: 1.3-1.5x1010
•Range: 0.9-3.5x1010
•Lead Time: 2 weeks
13. Normal Peripheral Blood
• NPB -> T Cells (untouched) isolation methods
• -/+ selection
• Fluorescence staining
- Trypan blue method
- AOPI method
• Cellometer method
Date
Tissue
Type/Unit
Lot Number
AO/PI Lot
Number
Cellometer
Count
Cellometer
Viability
Hemacytometer
Count
Trypan Blue
Viability
11-20-13
NPB-CD56+
5x10^6
A3860
130904-01
9.6x10^6
99%
10x10^6
96%
11-20-13
NPB-CD14+
10x10^6
A3871
130904-01
24x10^6
97%
22x10^6
91%
14. Custom Projects – AllCells Advantage
•
•
•
•
•
•
•
•
•
Custom tissue collection
Custom cell numbers
Custom processing/packaging
Custom aliquot and cryopreservation services
Custom cell based assays
Donor recruitment services
Multiple cell types from single donor
Donor-matched cells
Mobilized collections
16. Introduction to Nexcelom Bioscience, LLC
Innovation and Expertise in the Science of Cell Counting
• 10 years – design, manufacture and market cell enumeration and analysis
systems with image cytometry technologies
• Our approach: scientists help scientists to reduce error and increase assay
accuracy
• 50% employees with science degree, 8 field application scientists in US
• Work with lead labs in each research field to develop products
• Spread best practice to other labs
17. What is a heterogeneous cell sample?
How to select the right viability method?
Nexcelom Bioscience, LLC
Leo Chan, Ph.D. and Jean Qiu, Ph.D.
18. What is heterogeneous cell sample?
Heterogeneous cell sample contains target cells, other unwanted cell populations
and tissue debris.
Homogeneous
Purified CD34+ cells
Ficolled PBMCs
Whole blood
19. Develop better cell counting method for primary cells
• Dual fluorescence, nuclear acid binding AO/EB staining
protocol was developed at Stanford, Prof. Leonard
Herzenberg’s lab
(http://www.herzenberglab.org/Protocol-1/cell-preparation.html)
• NIH/Vaccine Research Center (VRC) used AO/EB & manual
counting
• Cellometer Vision automated AO/EB cell counting method for
NIH/VRC since 2009
• Nexcelom optimized AO/PI staining as the standard protocol
for primary cells
20. Dual fluorescence AO/PI viability assay
• AO is permeable to both live and dead cells
• AO binds to DNA and fluoresce bright green
• PI can only enter dead cells
- Binds to DNA of the dead cells
- Absorbs the green fluorescence of AO
- Produces bright orange / red color
• No signal is generated from non- nucleated
cells and debris
21. Leukapheresis samples:
over counting by bright field trypan blue method
Total cells: 119
Total FL cells: 39
Over counted cells: 80
They are not nucleated.
Over counting: 3x
22. The over counting ratio is sample dependent,
not a systematic trend
• Manual counting using Hemacytometer and trypan blue
• Automatic cell counting using Cellometer Vision and AO/PI
stains
• Experiment using Leukapheresis samples
Sample
Ratio
(Manual counting # to
Cellometer AO/PI #)
A
B
C
D
E
F
7X
5X
12X
3X
7X
5X
23. Heterogeneous cell samples mostly used
• Mouse splenocytes
• Human Peripheral Blood Mononuclear Cells
(PBMC)
• Human Mononuclear Cells (MNC)
• Human Leuko Pac
• Human Bone Marrow (BM)
• Human Cord Blood
• Mouse Tail Blood
• Bronchoalveolar lavage (BAL)
32. Using Trypan Blue Viability for Purified Primary Cells
AOPI stained all the CD34+ cells in BR
BR
AOPI
33. Which Cellometer is right for me?
• Trypan blue method is good for cell lines and purified primary
cell samples
- All Cellometer systems can perform Trypan Blue viability
measurement
• AOPI method is good for primary heterogeneous cell samples,
containing RBCs, platelets and cellular debris
- Cellometer systems with dual FL can perform AOPI viability
measurement
• Dual AOPI method was validated against total nucleated cell
counting method by lysing the RBCs
35. Benefit to Cellometer users
• Call: 978-327-5340 and you will be connected to a technical
support specialist
• Free in-lab training sessions (no limits)
• Free on-line support session via WebEx service
Nexcelom technical support specialist can connect to any Cellometer via
internet for training, trouble shooting and setting up new assays
• New in 2014: Monthly Cellometer Training Webinar
For new lab members and new applications
36. Q&A
Please use the prompt on the right of your screen to
send a question to the organizer
Questions will be read and answered
Any offline questions may be sent to
sales@allcells.com or sales@nexcelom.com
www.allcells.com/technical-support/
www.allcells.com/support/webinars/
www.nexcelom.com/technical-support/
www.nexcelom.com
Thank you!