I wish to look at gliosis in a dish, basically the neurons start becoming apoptotic and glial cells come to the rescue. But if I have a patient neurons, how do I mimic this event? Any idea what can be used a control?
By glia are you referring to astrocytes?
Depending on what your specific question is:
1. You could derive astrocytes from the the patient iPSCs or healthy control iPSCs.
2. You could seed these on a transwell so the cells don't interact but are signalling
or
3. You could seed them together and look at directly interacting effects.
Hi Yojet!
Without knowing precisely the question you're trying to answer, it may useful to look into glial toxin fluoroacetate in order to mimic gliosis.
Cheers,
Paul
Paul G. Morris Hi Paul, thank you for your reply!
I am trying to conceptualise an event in a dish (gliosis) where the neurons start becoming apoptotic and glial cells come to their rescue. A recent paper from Jennifer Lippincott-Schwartz's lab, where they showed using imaging that astrocytes protect the neurons from a toxic buildup. Which made me think, if that could be done in a phenomena of gliosis. Does it make sense?
Steven Fagan Hi Steven, thanks for replying!
Yes, by glial cells I meant astrocytes. Can you provide me a paper about the second point of transwell? That seems like more what I am looking for :)
Hi Yojet,
This paper uses a neuronal astrocyte co-culture transwell system well. Happy experimenting :)
Dear Yojet,
As you have found, "gliosis" has a huge amount of background literature. Both the CNS axon regeneration field and the excitotoxicity/stroke field have used the neuron/astrocyte co-culture model in the past to try to understand the role of astrocytes in neuroprotection, so I suggest looking at some of the older literature in those fields.
One of the original culture models for excitotoxicity with which I am familiar was developed by Dennis W. Choi, and used cortical neurons & astrocytes in contact with each other (there are other models). More recent Transwell studies were used to try to understand the role of direct contact vs. secreted factors in toxicity and rescue, as well as the inhibition or promotion of axon outgrowth. I also suggest looking at publications from Ben Barres' lab regarding astrocyte function and neuronal survival.
Transwells are a great approach. However, one system used to culture primary neurons used feeder layers of glial cells, with the neurons cultured on glass coverslips that were inverted over the glia. Since the distance of the coverslips can be varied by the size of the paraffin dots that support the coverslips, if you are already culturing your neurons on glass, you might consider that method as a first approach. Details of that method can be found in the book Culturing Neuronal Cells that is edited by Banker & Goslin:
http://cognet.mit.edu/book/culturing-nerve-cells
Good Luck!
Jill
Jill Miotke Hi Jill!
Oh sweet! The information supplemented is very useful! Thank you :)
Hello
Kindly look at links here
Article Aging in a Dish: iPSC-Derived and Directly Induced Neurons f...
https://fujifilmcdi.com/assets/CDI160913_iforumUS_neurotox.pdf
Good Luck
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