...nearly 145,000 cell lines. Cellosaurus Cellosaurus seeks to identify and describe all cell lines used in...in which the cell line was described. Using Cellosaurus Cellosaurus is searchable by cell line name (recommended...reached out to us to tell us about a resource called Cellosaurus, a growing cell line information database... related cell lines when annotating a cell line Community submissions via email: cellosaurus@sib.swiss...with cell line catalogs and collections. Importantly, Cellosaurus also identifies problematic cell lines...HeLa cells. Image courtesy of Josef Reischig.  Contributing to Cellosaurus Generated a new cell? Or ... about a cell line you’ve used? Submit your manuscript or data to Cellosaurus via cellosaurus@sib.swiss.This...

...Tracking lineage and cell identity at the single-cell level with CellTag CellTagging is a lentiviral based...individual cells. Each cell is labeled with 3-4 tags to increase the diversity of the CellTag library. ....    Find CellTag libraries here!  Applications of CellTag The Morris Lab developed CellTag to track the...reprogrammed cells based on single-cell gene expression patterns. The lab has also used CellTags for tagging...et al., 2019). Key takeaways of CellTag CellTag is a combinatory cell-indexing tool that combines scRNA-seq...describes the process where differentiated cells are turned into a cell type of choice while bypassing the intermediate...majority of cells failing to develop the desired identity. The development of single cell technologies...

...organs, cells comprise compartments and structures, called organelles. Take a sneak peak inside a cell with...vesicles via the secretory pathway to their subcellular or extracellular destination. At the end of their life...which are known to be a part of an organelle or involved in intracellular transport pathways, it is possible...proteins, however, in this case the cell needs to be fixed and the cell membrane permeabilized in order for... to reach their intracellular targets. Therefore, it can be beneficial to employ well-characterized marker...for labeling your subcellular structure of interest in the Allen Institute for Cell Science Plasmid Collection...and selected subcellular marker proteins, it’s possible to get an idea about the subcellular structures ...

...Chapter 9: Using stem cells as in-vitro models of disease. How Darrell is using stem cells from patients to... School. Darrell’s research focuses on the lungs, and, among other projects, using stem cells to develop...about Darrell, his research, and his focus on open science. Listen to the interview with Darrell Kotton...Introduction to Darrell Kotton and the Boston University Center for Regenerative Medicine. Darrell describes...him as a praticing physician and stem cell researcher. Darrell's ultimate goal - to develop therapies ...Kotton Lab. One of Darrell's favorite projects - differentiating surfactant producing cells that can be used...engineered cells in therapy. The lungs are difficult, but other tissues like the retina have cell-based therapies...

... horizontal cells, and Cre-dependent-GFP expresses in horizontal cells and ganglion cells, only the horizontal...improve cell-type specificity of AAV, work has focused on modifying the outside protein shell, or capsid... in retinal cells. To do this, the group developed a new strategy for finding retinal cell-type specific...and high cell-type specificity.  Strategy #3 (ProC): Create promoters by repeating mouse cell type specific... to test for cell type-specific expression of GFP. Half (113) expressed in retinal cells four weeks post-injection...targeted expression in specific cell types (ex: rods or cones) or cell classes (ex: photoreceptors which...occurred in one type of cell More than 50% of expression occurred in one type of cell, with the remaining ...

...ability to tell stories a similarly crucial skill for career advancement. Scientists do and should tell stories...interviews. They even tell short stories during weekly lab meetings. When scientists tell good stories, they...story short, telling a good story is something that can be learned, and will serve you well in your career...career. Story Telling at work Don’t believe me? Here’s an example of storytelling to answer an informational...be memorable. Work out what story you want to tell; tell it; stop. Many thanks to our guest blogger Jack...The part of my job I most enjoy is building and telling interesting stories — whether I’m working with ...when talking to friends or dates or colleagues we tell tales about parts of our lives that tend to have...

...more generally “How am I doing? Can you tell me what’s going well and what I could improve on?” Having ...have time to answer outstanding questions you have, well before experimental crunch time. Don’t: Wait to...at times! What makes it rewarding is when you can tell a trainee is understanding and things are ’clicking...

...quality work in cell culture. The problems with cell lines Simply put, the majority of cell line problems...HeLa cells, the first immortalized human cell line. He proved this by examining whether the cell lines...Cancer Cells, Marker-independent identification of glioma-initiating cells. The International Cell Line ...working cell bank has been made by a research institution (an excellent practice), or maybe cells have been...passage number, cell line, and date, which you complete each time you passage the cells (as well as recording...Animal Cell Culture Guide Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen Cell Culture... Outreach Scientist Nick Amiss. Cell culture in the present day Cell culture is a widespread tool used...

... to microbes by using non-invasive methods for cellular imaging with ultrasound. In his research he applies... take home message was that being creative and telling a story makes scientific topics both interesting...

...pursuit of achieving knowledge. Or, as we in the storytelling business like to say, science is all about navigating...all science communication, what you are doing is telling a (nonfiction) story. For there to be a story, ... journey? The same thing applies for science storytelling. You can introduce your results right at the...the science into. There are almost limitless storytelling options, and they all can be effective in the...circumstances.   However, in the midst of all these storytelling devices, we sometimes forget the most important...

...types of primary mouse cells, including neuronal and gut stem cells, dendritic cells, embryonic fibroblasts...fibroblasts, glia cells and neurons, and cell death post-transduction is low. Neuronal stem cells transduced... Primary cells recapitulate the natural biology of a cell type of interest better than immortalized lines...introduce a gene of interest into primary cells, so most primary cell lines require viral infection. A new ... primary cells. What if, instead of supplying a DNA construct, they could simply induce cells to take ...primary cell types using Cre and various loxP reporters. In both mouse and human embryonic stem cells, they...RJ, Rehmann H, Geijsen N. Cell. 2015 Apr 23;161(3):674-690. doi:10.1016/j.cell.2015.03.028. PubMed. Find...

...into 96-well plates, is extremely time- and resource-consumptive, since only wells with one cell can give...add a drop of serum to each well in the 48-well plate; rapidly growing cells such as HEK and CHO do not...while nearly all endocrine cells do. While not all of the wells in the 48-well plate will grow up, most ...Biosciences) to transition from a 48-well cell plate to a 96-well assay plate. We generally freeze at ... a wide variety of cell lines, from HEK to CHO cells to neural and endocrine cells such as AtT-20, PC12...then handled. Additionally, many cell lines, especially the endocrine cell lines we most commonly work with...and glucose. If we circled well-separated clones containing more than 50 cells (visible to the naked eye...

...psMEK1 in these same cells by 24-48 hours exposure to blue light also induced tail swelling in ~75% of worms...of Multiple Kinase Activities in Live Single Cells." Cell 157, no. 7 (2014): 1724-734. PubMed PMID: 24949979...Signal transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to ... dimeric Dronpa is less prone to aggregation in cells compared to the tetrameric Dronpa it was derived...inhibitors, psMEK1 and ERK KTR-mRuby2 expressing cells were incubated with an inhibitor of choice, exposed...inactivation, and then decreases. Treatment of the cells with a PP1/PP2A inhibitor leads to maintenance of... use rectal bacterial infection to induce tail swelling in C. elegans, but activation of Raf-MEK-ERK signaling...

...and analyzing autophagy of cytosol and organelles in yeast cells. Autophagy (Greek for “self-eating”) is...cytosol and organelles to the vacuole/lysosome for degradation and recycling. The key to Rosella’s autophagy-sensing...with the vacuole or lysosome. Rosella fluorescent properties Rosella is a dual color-emission biosensor...portion of Rosella: it will emit red fluorescence regardless of its localization in the cell. SEP’s pH ...color fluorescence Rosella emits based on its localization. Table 1: Summary of Rosella’s excitation and ...fluorescent properties of Rosella.   Table 2: Effect of Localization on Rosella’s fluorescence emission. ...Lysosome  ~4.8  Red    Using Rosella to study autophagy in yeast Rosella is targeted to the cytosol when...

...times), depending on the cell line so old cells should be retired! Conversely, cells younger than the 3rd ... Here are some tips for cell culture that will hopefully help you keep a well organized lab and contamination...hands! You should additionally remove all jewellery as well as that watch you’re wearing before you start...Make sure cells don't stay out too long While you’re preparing things, make sure your cells don’t stay...contaminants. Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine...’s best to split cells before the plates are more than 80-85% confluent because cells like their space...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red)...

...the SeqWell website to learn more about our plexWell kits and services! Many thanks to Joe Mellor, our...our guest writer from seqWell Inc. Joe Mellor is the Founder and CEO of seqWell, Inc. He is a sequencing... contributed by guest blogger Joe Mellor, Founder and CEO of seqWell Inc. Plasmids and PCR products are...DNA-barcoding step that uniquely labels DNA from each well of a 96-well plate. Then, samples from several plates are... to 1000s of samples individually, plexWell™ reduces every 96 well plate to a single tube after the first...across samples, NGS data obtained via plexWell™ libraries is well suited to large-scale de novo assemblies...from plexWell™ NGS data. Once Addgene produces plates of plasmid DNA, they are shipped to seqWell, where...

...Institute for Cell Science. Learn more by visiting the Allen Cell Explorer at allencell.org and the Allen...stem cell lines we’ve made using them are available as part of the Allen Cell Collection at Coriell. More... hiPS cell lines generated using these plasmids, please visit the Cell Catalog on the Allen Cell Explorer... tagging to elucidate cellular structures. Overexpression methods flood the cell with protein, which can... in human cell lines. Furthermore, the quality control necessary to ensure the edited cells are behaving... and images generated from these cell lines can be found in the Cell Catalog on our data portal. The plasmids...

...varying across cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher... of your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle, when...HDR events, ensure cells are actively cycling through the cell cycle stages. If cells are overly confluent...way to make a knock-in cell line is to utilize the built-in repair pathway cells already have for repairing...than quiescent cells. It is important to know that HR is often mutated in cancer cells, specifically in...keeping an eye on those cells! To take this one step further, you can inhibit cell cycle regulatory proteins...negative outcomes on viability if cells are arrested too long. Prolonged cell cycle arrest can lead to apoptosis...

...that already do mammalian cell culture. HEK-293T cells are the most common cells used in viral production...versus insect cells for viral production Consideration Mammalian cells Insect cells Viruses AAV ...type of cells that will be used to make the virus. Viruses can be produced in many different cell types,...produced in bacteria cells. However, some viruses can be produced in multiple different cell types, including...you make an informed decision. Mammalian cells Mammalian cells are a great option for viral production,...production in mammalian cells is relatively quick. After transfection, HEK-293T cells start producing virus...using qPCR or digital droplet PCR. Insect cells Insect cells are a less common option for the generation...

...prevent stem cell lines from becoming the new HeLa cells?), genome engineering and stem cells, and the difficulty... Podcast Interview with Stem Cell Researcher Darrell Kotton 2. Stem cell scientists care about patients... landmark Cell paper showing that it was possible to generate induced pluripotent stem cells (iPSCs) capable...forming nearly all cell types in the body from otherwise terminally differentiated cells simply by expressing... happening. Stem cells scientists have certainly not lost site of that. 3. Stem cell science is hard, ...sharing of his OSKM factors have affected the stem cell community – a community largely created due to Yamanaka...understatement. Multiple speakers from top minds in the stem cell field have described the Yamanaka paper as "life...

... do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were transferred...functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside the...a wide range of cell types including but not limited to muscle, liver, and brain cells. There are a multitude...chain (c) and is characterized by a lack of immune cell development and function. To date, gene therapy ...that can deliver gene therapies directly to the cells that need them. Adeno-associated virus (AAV) – based...delivery of nucleic acid cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery... delivery vehicles with great safety profiles as well as other benefits. What makes AAV vectors great...

...single genetically-defined subset of cells. By labeling fewer cells in a population, it’s easier to visualize...non-overlapping cells. While transgenic animals are commonly used to specifically manipulate a cell type of interest...expressed. In cells where the stop cassette remains, the effector is not expressed and the cells are not labeled...sparse labeling of the cell population that expresses SPARC and the fraction of cells that will be labeled... interest in an organism, all cells of that type are affected. Previous tools to overcome this restraint... of the gene construct. A predictable number of cells will express the effector or retain the stop cassette...tool that allows for the labeling of a subset of cells with the same genetic makeup, overcomes all of these...

...hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells can be implanted...Annotation of genes in immune cells typically involves the creation of germline knockout mice, which ...CRISPR-based systems enable gene knockout in immune cells in a high-throughput manner, but these systems have..., ubiquitous knockout of single genes in immune cells in vivo (LaFleur et al., 2019a and LaFleur et al...many genes have different functions in different cell types, at different times, and in combination with... L-CHIME Knocks out a single gene in defined cell types (lineage-based) pXPR_070 Violet-excited...benchmarked on positive control genes in CD4+ and CD8+ T cells. To demonstrate the power of these systems to investigate...

...HEK293 cell culture and in live zebrafish. For cell culture experiments, only dL5**-expressing cells treated...inactivate target proteins or cells while having little effect on neighboring cells. The photosensitizer used...Here Light-induced cell ablation with MG-2I-dL5** He et al tested out light-induced cell ablation with MG...effective killing of dL5** cells without death of neighboring bystander wild-type cells.   Zebrafish experiments...fewer mCer3-labeled cells in their hearts 24 hours post treatment and no mCer3 cells present 96 hours post...increase cell ablation efficiency, but dL5** phototoxocity is induced when expressed on the cell surface...protein inactivation/cell ablation while having limited collateral damage to bystander cells. Are you ready ...

...never considered the sex of their cells. For one thing, cells and cell lines degenerate over time—and sex... effects of using all female cells and tissues, specifically in stem cell therapies. Let’s go back to ...therapeutic capacity of stem cells. Yet, very few researchers consider the sex of their cells when doing experiments—which...sexes in basic research is excellent design), but they used all female stem cells—this was an unconscious... stem cells. Nor did they detect important differences in function between male and female cells. The ...experiment. In the case of stem cells, these factors may include cell type, the disease being treated,...sex of cells and tissues—when that information is reliable—is important to ensuring excellence in basic...

...sequence change the cell must undergo homology-directed repair (HDR), a distinct cellular DNA repair pathway...incorporated into the edited genome. In mammalian cells such as stem cells, HDR is relatively rare and DSBs are predominantly...them into somatic cells affected by the disease and study what’s wrong with the cells. However, our excitement...Munich and Marc Tessier-Lavigne’s lab at the Rockefeller University in NYC. The CRISPR/Cas9 system is ...sequence-specific changes in human induced pluripotent stem cells (iPSCs). The CRISPR/Cas9 com­plex is very efficient... stranded breaks (DSBs) into genomic DNA in many cell types and often results in biallelic modifications... After genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al, Mai et al) we...

...gain access to some cells and tissues, such as human induced pluripotent stem cells (hiPSCs). Nanoblades... into particles at the cell membrane. Mature Nanoblades then bud into the cell culture supernatant and... Addgene. Genome editing primary cells with Nanoblades Primary cells are often refractory to gene delivery...mouse bone marrow (BM) cells, primary human hepatocytes, and hematopoietic stem cells (HSCs) using Nanoblades...-70% for these four cell types (Table 1). Additionally, hiPSCs and mouse BM cells still behaved like iPSCs...Primary cell type Gene targeted by guide Editing efficiency hiPSCs  EMX1  67% Mouse BM cells from GFP...mouse BM cells when using a gRNA targeting the Fto gene, while editing was not detected in cells electroporated...

...CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via FACS, primer validation...article "Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9" from the Journal of Visualized...CRISPR/Cas9 to create genomic deletions in mammalian cell lines. Below Bauer and Canver discuss the motivations...deletions is to produce monoallelic deletion clones. Cells with just a single intact copy at a given locus ...screening by conventional PCR. Biallelically disrupted cells can be easily identified. Even in the case of clones...methods for CRISPR design, cloning, and delivery, as well as their techniques for creating deletions, identifying... His lab studies the molecular genetics of blood cell development and disease. Learn more at his webpage...

...help in the capture of other rare cell types, like stem cells or cells infected with malaria or other parasites...capture cancerous cells expressing low levels of tumor antigen. I asked Howarth to tell us more about SpyLigase...looking at, magnetic isolation of circulating tumor cells, we found that the binding stability really mattered... in the pili that they use to anchor on to host cells. Addgene: Were there any major hurdles along the... donor for keeping going while we worked out the cell capture system.) Cancer and other applications Addgene...cancer? Howarth: The capture of circulating tumor cells (CTCs) is going to have a big impact on our understanding...Addgene. Howarth: We submit all our plasmids that may well be of use to others to Addgene. I am a big fan; ...

... competent cells (protocol)  Transformation of electrocompetent cells (protocol)  Cells prepared using...plasmid that transforms well in other competent cells) to ensure that your cells can be readily transformed...to make your own competent cells It’s easy to make your own competent cells. If you need a high efficiency...purchasing competent cells. However, for most everyday cloning, homemade competent cells will work just fine...Preparing chemically competent cells (protocol)  Preparing electrocompetent cells (protocol)  Untergasser...of chemically competent cells (protocol) Preparation of electrocompetent cells (protocol) At Addgene...Set  from Zymo Research to make competent cells because cells prepared using this kit can be transformed...