Hello everybody.
I'm trying to culture and differentiate murine DPSC into neurons like cells, but the proliferation ratio is really low.
Moreover, present cells show different morphologies (no genetic analysis done yet).
Do you know any coating for the plate that would increase proliferation and differentiation into neurons like cells?
Thanks ;)
Francesco, what have you tried for coatings? I never have worked with DPSCs, but have made human and murine neurons on gelatin, laminin, and matrigel. Have you tried any of these? Are you certain that it is the coating, rather than some other aspect of your protocol, that is at fault?
You can try coating with Laminin-5, it has a positive effect on DPSCs proliferation.
Hello Francesco,
First, Proliferation and differentiation are two contradictory things...if u want to do efficient differentiation then u have to stop cell proliferation so that cells can differentiate properly. Second, coating is done to increase adherence and sometimes Fibronectin and laminin help in increased neural differentiation as they help in Na/K channel activity and hence increased survival and differentiation of neurons.
As you cell proliferation is slow, so I will suggest you to standardise an optimum media first (preferably alpha-MEM works best for DPSCs+ good quality FBS), once you get desired proliferation upon standardised conditions then you can try differentiation without coating and even with coating parallely (FN and laminin). As per my experience with human DPSCs, I get very efficient neural differentiation even without coating.
Hope it helps !
I agree with Ajay. First you must try to standarize your DPSC culture conditions in order to get enough cells for you differentiation protocol. When trying to differentiate into neurons, cells become spindle with less cytoplasm area and deveolpment of projections. This is when you need to increase cell adherence. In my expercience, differentiation protocols must be made over coated dishes (poly-lysine for example) and a large amount of cells, considering that at least one third of the population would detach.