Currently I am doing Low density hippocampal cultures. The neurons are growing nicely, and with very little cell debris.The problem that I am facing is that they are getting easily contaminated. I have tried all troubleshoots but nothing seems to workout properly. Is there anyway to save my cultures?
What type of contamination are you getting, mold, yeast or bacteria?
When is the last time that your cell culture hood was cleaned? Also, tissue culture incubators needs to taken apart every six months or so and the removable parts autoclaved if possible. Molds and spores can be seasonal and will build up in the incubator. You can see this if you wipe the surface with a towel, it will be blackish, grey.
Primary hippocampal culture are highly sensitive to contamination in comparison to basic cell line culture (medium and component like B27 seems to be very well appreciate by bacteria and fungi). Adding antibiotic is one aspect but not enough. You need to use sterile tools (Forceps, dish, cover slip....) at every step of your procedure. Another aspect is, if you are sharing space, tools, media, components (Hood, Incubator) with line cell culture users, they might have no contamination but you do because incubator and hood are contaminated ( as Jonathan mentioned).
My opinion: clean carefully absolutely everything from your working space and start with fresh new media.
Good luck!
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