I any designing a microwell assay to measure spheroid forming potency of neural stem cells on a single cell basis. I could not find any papers that specifically listed whether it is or is not possible.
Neural stem cells harbor the self-renewal potential which is responsible for the formation of neurospheres. There is a reversible change between neural stem cells and progenitor cells. As they proliferate gradually, they begin to secrete neural growth factors p75, hepatocyte growth factors etc.
theoretically yes... but most ideas on this are correlative. However depending on what stage you harvest your progenitors you will get very different 'types' of spheres. You should be able to test your single cell question in your microwell if you also have access to a good imaging system to track multiple individual cells in, say, a 96 well plate or microwell system
If I am remembering correctly that assay is described in the original neurosphere paper from Reynolds and Weiss. You plate your stem cells in a 96 well plate with a concentration of 0.2 cells per well to make sure that there is only a single cell per well and then determine the potency of your stem cells by how many spheres are growing in your wells.
Hi
hope this article could be helpful:
Methods Mol Biol. 2008;438:173-84. doi: 10.1007/978-1-59745-133-8_14.
Clonal analyses and cryopreservation of neural stem cell cultures.
Gritti A., Galli R, Vescovi AL.
The answer is yes, they can, certainly for embryonic NSCs. I've done this myself, plated at 0.5 cells per well. You do need a lot of patience, as they grow very slowly to start off with in these conditions. Also, plate plenty more wells than you need, as many won't have the correct number of cells in them, and many others that do won't grow. Never tried it with adult ones, I would suspect that they are trickier to do this with.
Hi Niels. How long does your cells take to form spheroid and what are your respective sizes? For me the sizes depending on the stem/progenitor cells. My cells ranged from 50-300 microns
Sizes sounds about right, especially for embryonic NSCs. As for timings, to be honest, it's been a few years since I've done it, so I don't exactly remember, but I think about three weeks at least.
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