...Pre-implantation Blastocyst Cell Types from Fibroblasts. Cell Stem Cell. 2019 Jun 6. Buganim Glial Cells Retinal Ganglion... Plasmids Stem Cell Research Plasmids Stem cells are proliferative, unspecialized cells with the capacity...different cell types in the body during development. Unlike other cell types, each stem cell has the potential...generate different cell lineages. Pluripotent stem cells, such as embryonic cells, can potentially give...any other cell type in the body. Multipotent stem cells, such as adult (or somatic) stem cells, can differentiate...disease-specific cells. For this reason, iPSCs have become an excellent unlimited stem cell source for therapeutic... of Stem Cell Research Plasmids . For more information on stem cells, visit the NIH Stem Cell information...

... Depositor Collections Cell Migration Consortium (CMC) Plasmids Cell Migration Consortium (CMC) Plasmids...collaboration between the Cell Migration Consortium and Addgene. Member laboratories of the Cell Migration Consortium...This resource is a collaboration between the Cell Migration Consortium (CMC) and Addgene, which highlights...Consortium have deposited plasmids useful for cell migration research at Addgene. CMC investigators who have...MIT Rick Horwitz , University of Virginia Denise Montell , University of California, Santa Barbara Martin...

...obtain approximately 30 cells per 96-well plate. Include 100 μl per well of cell culture media. For the...is for suspension cells. Adherent cells can either grow as individual cells in 96-well flat bottom plate... While MEL cells are suspension cells, instructions for adherent cells have also been included. Ensure... Fluorescence Activated Cell Sorting (FACS) of Transfected Cells Prepare cells for FACS by filtering them...limiting dilution at 30 cells per 96-well round-bottom plate. Optimize plating the cell type used at limiting...parental and bulk sorted cells by resuspending parental and bulk cell pellets in 50 μl of DNA extraction...suspension cells, transfer all clones to a single 96-well plate that already contains 50 μl cell culture ...

...Consortium plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...expression plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...screens or cellular assays to identify novel interactions or pathways that are important in cellular recognition...AVEXIS (for Avidity-based extracellular interaction screen) in which the extracellular regions of receptor ...and Wright GJ Molecular & cellular proteomics 2010 Dec; 9(12):2654-65. A cell surface interaction network...screening for novel low-affinity extracellular protein interactions. Bushell KM, Söllner C, Schuster-Boeckler... Plasmid Collections EXPRESs Plasmids EXtracellular Plasmid RESource (EXPRESs) Consortium You may...

...give the cell its shape, organize cellular organelles, and are necessary for movement and cell division...these well-characterized proteins. General Structures The cell contains a variety of organelles and subcellular...between animal cells. They are important for holding cell structures together and reducing cell stress. There...Collections Fluorescent Proteins Organelle Markers Fluorescent Proteins: Subcellular Localization More Fluorescent...proteins that are targeted to various organelles or subcellular structures. You should fuse your protein...example, houses the cell’s genetic material. The mitochondria produces the energy cells required for chemical...the table below to find markers of general cellular organelles and structures. ID Plasmid Structure Targeting...

...for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key cellular structures... Protein Guide Subcellular Localization Plasmids Protocols The Allen Institute for Cell Science has produced...fluorescently tagged proteins of key cellular structures in human cell lines of their choice. For a full ...Tagging in Human Cells on the Addgene Blog. Plasmids for Endogenous Gene Tagging in Human Cells Find additional...additional cell line information from the Allen Institute for Cell Science data portal (Link opens in a new...images from the hiPS cell lines gRNA sequences used in the creation of the hiPS cell lines The following...plasmids can be used to create cell lines with endogenously-tagged cellular structures. Addgene ID Plasmid...

... sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to living cells To learn...repair protein, and is suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag ... (CoA). These tags are particularly well suited for labeling cell-surface proteins, and are useful for...Control for cell surface localization 101131 pACP-GPI Control Plasmid ACP/MCP-tag Control for cell surface...visualization of proteins inside cells. Depositor... Depositor Collections New England Biolabs Cell-Imaging Plasmid Collection...Collection New England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental Information ...

...media and cells. Centrifuge at 3900 rpm for 20 min at 4 °C to pellet the cells. Keep the cell pellet on ice...Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment...resuspend each pellet completely. Combine resuspended pellets and keep on ice. Process the cell pellet from above... HEK293T cells is critical for optimal AAV yield. Do not overgrow your cells. Pass the cells twice a week...multiple times to obtain a single cell suspension (no clumps of cells). Pool cells from 2 x T-175 flasks. Adjust...obtain a single cell suspension (no clumps). Count cells using a hemocytometer or cell counter. Seed 350...Harvest cells and media by tapping the sides of the CS5. Cells should detach easily. Transfer cells and media...

...control well). Perform a "reverse transduction" by seeding 50,000 cells into each well of the 6-well dish.... Aliquot 1 mL of cell suspension (i.e., 50,000 cells) into each well of the 6-well dish. This brings ... the cell media. Cell death by some cells in the culture may adversely affect the surviving cells in the...single viral dilution to each well (each well gets one dilution, so a 6-well plate will hold 5 dilutions...media to make the cell solution in this step. To seed the cells: Prepare a batch of cells as follows: Dilute...stable cell pool. Observe the dish every day to ensure that the cells in the untransduced well (0 µL lentivirus...conditions for the surviving cells. Even in the absence of cell death, the cell media should be changed every...

...0.5 cells/well. Seeding an average of 0.5 cells/well ensures that some wells receive a single cell, while...Seed individual cells in a 96-well plate Day 2–14: Monitor cells for growth and expand cells Day 14–30: Analyze...homogenized cell solution. Homogenized cell solution concentration: 4 × 10 5 cells/mL To seed one 96-well plate...of a 5 cell/mL solution. Calculate the total cells needed: Total cells needed: 10 mL × 5 cells/mL = 50...any well receives more than one cell. Pro-Tip In each 96-well plate, seed one of the corner wells with...within each well, especially in the corner of the well as cells tend to settle there. Once the cells have expanded...generate a monoclonal cell line from a polyclonal pool of stable cells. Transducing cells with lentivirus results...

...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 ...

... 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 ...

...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...

...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...

...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...

...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...

... 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...