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help on cell type identification from mouse EBs

huge cells beating cells

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#1 DavNu

DavNu

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Posted 27 March 2012 - 07:08 AM

hello

I did EB formation from mouse ESC and plated them on gelatin. I then incubated it with a proneural minimal medium (n2 supplement essentially dmem f12 glutamax). I got two or three per well and after plated EB expension for 10days I got cells emerging from an EB (still compact) but the cells migrate in a circle motion creating a comet tail structure ( pict1). Cells within this structure are long narrow and slim and really elongated those cells were with no motion but after few days some those cells are beating with a high rate almost constantly (pict2). I previously worked in cardiogenesis from ES and beating area in EB arise from compact patches of cells creating a continium of actinin positive cells. Here those cells beat independanlty and heracticly. It must be muscle cells but it is strange for cardiomyoctes and for muscle it does not fit with the pacemaker activity.
Thus I will be happy to hear your suggestion ( I can not do immunofluorescence yet on them and if with what?)
here is pictures of the cells.

1 eb bj1.jpg

zoomed

2 eb bj1+.jpg


This was type1 of unknown cell type then I have a second type of unkown cells in the same well.
Those cells I really do not have a clue what it can be. It is still in N2 condition. the cells are huge with a defined shape and a clear nuclear menbrane.
it seems that there is two type of cells one well spread and one strongly refringent and round.

type2b.jpg


another zoomed

type2.jpg

the cell seems to contains something but it doesn't look apoptotic autophagic or senescent.

Ebs can make almost any cell type but in my experience of EBs I never saw something like that and
If any of you have an idea of the cell type of those cells thanks you for your anwsers.

This is just out of pure curiosity and not something i am struggling with.

best regards

Attached Thumbnails

  • 0002.jpg






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