To keep your PSCs in a pluripotent state it is important to handle them with care. PSCs require strict culture media and passaging to keep them healthy and undifferentiated. The two culture systems commonly used for PSCs are either feeder-dependent or feeder-free culture systems.
Feeder-dependent cultures use a layer of feeder cells which will provide the PSCs with the growth factors and extracellular matrix proteins they need to keep them healthy and able to expand. MEFs are commonly used as feeder cells.
Feeder-free culture systems use replacement media which provide all the growth factors the PSCs need. This may make things much easier as the cells can grow on simple matrix-coated plates, so you won’t need to worry about removing the feeder-cells for downstream experiments. You can also use biochemicals to help maintain pluripotency in feeder-free culture such as SC-1 (Pluripotin), Dual kinase and GTPase inhibitor.
It is also important to check that your PSCs are still in a pluripotent state. A good way to check for this is to use undifferentiated cell markers in applications such as western blot or ICC. Presence of undifferentiated cell markers such as Oct4 and Sox2 are a good first step to check if your cells are pluripotent.
We recommend that you use these undifferentiated cell markers:
You could also try our embryonic stem cell marker panel, to find the right marker for your research.
Undifferentiated cell markers like those listed here are expressed by undifferentiated PSC however using these markers alone does not guarantee that the cells you are working with are pluripotent. To be entirely certain of pluripotency it is essential to also check your PSCs using a functional assay.