How can I sort human neural stem cells without killing them?

I tried to sort human neural stem cells (induced from hiPSC) and observed that almost all of the cells die after the sort, although they were alive before. I used 100um nozzle, and PBS-EB (EDTA...

31 December 2015 5,851 7 View

Fluorescent signal reduction after fixation/permeabilization?

I infected human neural stem cells with a RFP tagged viral construct, and wanted to sort infected and undifferentiated cells. For this I use two intracellular markers, Sox2 and Nestin. When I ran...

31 December 2015 6,499 7 View

Any advice on a FACS analysis of iPSC?

I tried sorting human iPSC by flow cytometry, but lost a lot of cells during staining or transfer of cells from tubes to epis or so. Any suggestions to prevent this? I know that some people coat...

11 December 2015 8,727 4 View

Has anyone performed LTC-IC assay on human CD34+ HSC with hTERT immortalised mesencyhmal stem cell line, SCP-1?

02 March 2015 4,047 1 View

Neurosphere like formation from human NSC?

I derived human neural stem/progenitor cells from iPSC and have been doing RNAi screen on the NSPC. Normally they are adherent cells and form a network like structure. But when I knock down this...

01 January 1970 7,801 5 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

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28 February 2021 9,936 2 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View