trouble with primary adult neurosphere cultures - (Oct/05/2007 )
I have been working on primary neurosphere cultures from adult mice for over 3 years with good results (protocol as from Doetsch et al.). A few months ago I suddenly started having serious problems, with a progressively low yield of spheres up to having no spheres at all. There was nothing new in the protocol I could relate these problems to, and since then I have tried everything (new reagents, new medium, new protocols!) but no results. Does anybody have any clue on what might have happened?? Also, a termometer in my incubator indicates a 38°C temperature instead of 37° as indicated by the display: could this have something to do with such a mysterious loss of neurospheres?!
Ah, the joys of primary culture . I have not worked with neurospheres, but the fact that your thermometer reads differently from the display could be a warning sign of incubator trouble: you may have to check the CO2 level, and the humidity.