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We narrowed to 820 results for: cell

Showing: 51 - 60 of 820 results

  1. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...Pluripotent Stem Cells from Blood Cells: An Efficient Protocol Using Serial Plating of Reprogrammed Cells by Centrifugation...human-induced pluripotent stem cells from cord blood and peripheral blood cells. Stem Cells, 2013. 31(3): p. 458...possible to reprogram differentiated cells into induced pluripotent stem cells (iPSCs) by the addition of reprogramming... somatic cells that are otherwise difficult to obtain. These can then be used to perform cell therapy ...in vitro. Find stem cell plasmids at Addgene Many sources of adult somatic cells have been probed for...Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic...stem cells from a small amount of human peripheral blood using a combination of activated T cells and ...

  2. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...performed on whole-cell extracts from the initial stable cell pool (pool) and from individual cell expansions ... in several human cancers) in A549 cells (a human lung cancer cell line), armed only with viruses obtained...need our A549 cells to express Cas9. In these particular experiments, we generated A549 cells stably expressing...52962-LV, Figure 1). Some cells were then used to generate a monoclonal cell lines by limiting dilution...The rest of the cells were frozen down. Figure 1: Cas9 expression in A549 cells transduced with...blotting was performed on whole-cell extracts from stable pools of A549 cells transduced with Cas9 expressing...A549 cells are relatively easy to transduce. I ended up with too many Cas9 integrations per cell, and ...

  3. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ... 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...protocol is for suspension cells. Adherent cells can either grow as individual cells in 96-well flat bottom...Deletions in Mammalian Cell Lines CRISPR: Protocol for Genomic Deletions in Mammalian Cell Lines Other Addgene...detail for murine erythroleukemia (MEL) cells, a suspension cell line. The culture medium in all steps ...numerous cell types using preferred culture conditions and transfection strategies for each cell type. While...Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation solution...

  4. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    ...prevent stem cell lines from becoming the new HeLa cells?), genome engineering and stem cells, and the difficulty... 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... Podcast Interview with Stem Cell Researcher Darrell Kotton 2. Stem cell scientists care about patients... 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...

  5. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ... 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...specific cell line, and its transfection efficiency. We often trypsinize the transfected cells on the day...increased). For some cells, add a drop of serum to aid in outgrowth. Depending on the cell type, we sometimes...growing cells such as HEK and CHO do not seem to require this, while nearly all endocrine cells do. While...difficulty deriving single clones from SH-SY5Y cells, as these cells did not seem to form discrete colonies but... of Medicine. In the Lindberg Lab we often make cell lines that overexpress genes of interest; more recently...

  6. AAV for Neuronal Tracing

    Type
    Collection
    ...spread from this cell to synaptically connected cells (meaning, it can spread to cells that are projecting...infect a single "starter" cell, but new virus cannot spread to any neighboring cells. To enable monosynaptic...provided to the starter cell in trans , via delivery by an AAV. Since this starter cell now has all the viral...projecting to the starter cell). Once in the secondarily infected cells that lack the G protein, the virus ...connected cells. To further refine the targeting of G-deleted rabies to specific starter cells, the tropism...G-protein is delivered to the starter cell. (2) The starter cell now expresses TVA and G-protein. EnvA-pseudotyped...is delivered to the starter cell, and specifically targets the starter cell via the EnvA receptor. RABV...

  7. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    ...model systems: either human cells or mice. In a recent issue of Cell Stem Cell, Musunuru, Chad Cowan and...done in stem cells, we can differentiate those mutant and wild type stem cells into any cell type relevant...system for generating human stem cell-based disease models." Cell Stem Cell. 12, 238-251 (7 February 2013...homology-directed repair by flooding the cell with a mutant template and fooling the cell into using that template for...mutations influence the disease process. In the Cell Stem Cell paper, for example, we generated hepatocytes...generating the mutant cell lines that one needs to do disease modeling in stem cells. Addgene: Tell me about...Kiran Musunuru's lab in Harvard's Department of Stem Cell and Regenerative Biology is to understand the basis...

  8. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    ... into cells in the lab. The introduction of foreign DNA or RNA into bacteria or eukaryotic cells is a ...application of an electrical charge to cells, to increase cell membrane permeability and thus transformation...introduce foreign DNA into eukaryotic cells, like mammalian cell lines. This can be done with lentiviral...the genome and stable cell lines, where foreign DNA is incorporated into the cell’s genome and is thus passed... pokes holes in the cell membrane so DNA can be introduced directly into the cell. Microinjection requires...multiple ways foreign DNA can be introduced into cells including transformation, transduction, conjugation...genetic material from the environment by bacterial cells. In nature, this genetic material often comes from...

  9. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    ...human embryonic stem cells. Transduced stem cells were differentiated and single cell RNA-seq was performed...transduced human embryonic stem cells with the viral TF library, with each cell receiving only 1 TF expression...construct. Cells were cultured following a differentiation protocol before performing single-cell RNA sequencing...differentiation.   Fig. 2: Single cell RNA-seq clustering of cells transduced with the TF pooled library... clusters of individual cells along with color coded nominated reference cell types.  (Adapted from Joung...delivered to each cell, future applications could deliver multiple TFs to each cell, allowing for combinatorial...generate unique cell types. Of the 24 TFs they tested, 22 expressed markers consistent with the cell type that...

  10. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...Love your cells and they'll love you back A successful lentiviral infection begins with your cells, both ...both your packaging cells and your target cells. Poor tissue culture practices can lead to low titers and... promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...reaching 108 cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients...consumption due to the high cell density of the culture. In addition to benefitting your cells, medium exchanges...therefore titers obtained in one cell type may not translate well to other cell types. To overcome this issue...
Showing: 51 - 60 of 820 results