mouse spleen cell culture - (Aug/19/2006 )

Hi,

I am trying to see G2/M arrest in mouse spleen cells. I use RPMI 1640 with L-glutamine & hepes & 10% Fetal calf serum as medium. I see lot of cell death even in control cultures . Can some one pls. sugest a way to improve the culture conditions.

Hi rnj,
we also using same things but you here not telling about all procedure you using for its, i mean from isolation , single cells preparation and plating in detaile. In our experiment we are using pickup spleen and crush betwwen two rough surface slides by adding incomplete media and keep foe some time depbrss settle down then transfer supernatent in other 15 falcon tube and spun its.
Now you get a pellets of white and red cells now you needed RBCs lysis which done by using 10X PBS in 100ul in disspersed pellets and after 10sec. add 900ul of triple disstiled water now againg added 5ml media and spun its this steps repeat till you get pellets of white cells now finally cells dispersed in RPMI complete media with 10% FBS or FCS and 1X antimycotic and antibiotic in complete media and plate its according to their experiment in culture plate.
I thinks this way you able to minimising cells death.
all the best and good luck.
you can also contact me directaly via mail my id is awadh507@gmail.com

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