21
$\begingroup$

Here are some pictures of human iPS cells in culture. After several passages, some cells look really weird, can you help me understand what is going on? What am I looking at? What are those blastocyst-like structures? (bluish picture) and what about that cell full of vacuole-like thing?

P.S. do not pay attention to the scale bar, it's incorrect.

enter image description here

enter image description here

enter image description here

enter image description here

Extra info:

I did not induce them, I got them already dedifferentiated so I can't be specific on that part of the protocol. However, I am growing them in Essential 8 Flex Medium (with supplements as specified from the kit protocol), I add rock inhibitor 4h before passaging them. Passages are done by dissociating colonies using EDTA only (no enzymes). Most of the expanded cultures look normal, some showed the formation of embryonic bodies (the last picture added) but the blastocysts are very different and not really expected.

Improve this question
$\endgroup$
11
  • 1
    $\begingroup$ I am not entirely sure but there is some vacuole formation so maybe they are going towards the apoptosis. However to see clear phase contrast between Feeder layer and iPS cells the light diffraction may give you hallow formation around the radius of the cells. (Source: Dad) $\endgroup$
    – cheesebread
    Commented Sep 21, 2016 at 15:06
  • 1
    $\begingroup$ What further changes on them happened later-on? $\endgroup$
    – user25568
    Commented Sep 22, 2016 at 18:44
  • 1
    $\begingroup$ This question would be easier to give an answer to if you provided specifics on your protocol for inducing and culturing iPS cells, any variations from typical protocols, any experimental treatments, etc. The greatly elevated vacuole formation suggests cytotoxic stress. Elevated autophagy can be cytoprotective if the cell is moderately stressed, but it is a precursor to apoptosis when the stress cannot be alleviated by autophagy. Depending on the scale (please clarify), the blue picture may depict an "embryoid body" (essentially a blastocyst) - see "identity" section in your own wiki link. $\endgroup$
    – Demosthenes' pars triangularis
    Commented Sep 22, 2016 at 20:28
  • 1
    $\begingroup$ I did not induce them, I got them already dedifferentiated so I can't be specific on that part of the protocol. However, I am growing them in Essential 8 Flex Medium (with supplements as the kit specified), I add rock inhibitor 4h before passaging them. Passages are done by dissociating colonies using EDTA only (no enzymes). Most of the expanded cultures look normal, some showed the formation of embryonic bodies (the last picture added) but the blastocysts are very different and not really expected. $\endgroup$
    – alec_djinn
    Commented Sep 23, 2016 at 9:59
  • 2
    $\begingroup$ @alec_djinn Just a couple examples, though I may be mistaken. (ref) (ref) $\endgroup$
    – CKM
    Commented Jun 4, 2017 at 0:12

1 Answer 1

Reset to default
4
+50
$\begingroup$

they might be embryoid bodies:

https://en.m.wikipedia.org/wiki/Embryoid_body

Improve this answer
$\endgroup$
2
  • 1
    $\begingroup$ looks like it, at least in the last picture. $\endgroup$
    – alec_djinn
    Commented Sep 5, 2017 at 7:00
  • $\begingroup$ ips cells fascinate me to no end. I wish I could culture them at home. $\endgroup$
    – 4D Neuron
    Commented Sep 5, 2017 at 22:52

You must log in to answer this question.

Not the answer you're looking for? Browse other questions tagged .