Hi every one, we have extracted neural stem cell by neurosphere assay from adult 6 week old male mouses. after 5 -7 days we had some neurosphere like structures, we also had very much debris then we passage them on day 7 but we lost lot of them. Also I don't know which day I should renew the mitogens. which size of neurosphere is the optimum size for nerosphere to be passaged?
I was Interested some one give some advises about the best condition I should hold after extraction?
Thank you
Havva Mehralitabar
Hi,
check this publication
Optimal time for passaging neurospheres based on primary neural stem cell cultures by Fangling Xiong et al. (Cytotechnology. 2011 Dec; 63(6): 621–631.)
hi
I think you must change some of your culture condition:
1) first of all change your NSC source and use neonatal mice or pregnant mice
2) for culture media use N2 supplemented media (cat; AR003) or NSC basal media ( cat; SCM003) and change media each 2 to 3 days
3) for norospher size selection some researchers use 50 to 100 micron spheres
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