2 replies to this topic

[mia]

Hi! Could anyone provide information on how to culture neuro2A and neuroblastoma B104 cell lines? Do these cells need a particular coating to grow on? What medium do they prefere- I read about DMEM + 10 %FCS (or, after plating N2 supplement), but does anyone have experience with the use of RPMI medium + 10% FCS?

[klinmed]

mia, on May 31 2009, 02:11 PM, said:

Hi! Could anyone provide information on how to culture neuro2A and neuroblastoma B104 cell lines? Do these cells need a particular coating to grow on? What medium do they prefere- I read about DMEM + 10 %FCS (or, after plating N2 supplement), but does anyone have experience with the use of RPMI medium + 10% FCS?

Neuroblastoma cell lines are usually very easy to culture. These cells will grow well on standard tissue culture plastic (absolutely no need for coated surfaces).  In the past I routinely used RPMI/10% FCS + 1mM Na pyruvate for human neuroblastomas like  BE2©. It should work fine in your hands.
Harvest confluent cultures using trypsin but remember not to over split. Would suggest 1:2 - 1:3 for neuro2A. Can split B104 using higher ratios (eg 1:5).

Hope this helps.

[mlk]

mia, on May 31 2009, 05:11 AM, said:

Hi! Could anyone provide information on how to culture neuro2A and neuroblastoma B104 cell lines? Do these cells need a particular coating to grow on? What medium do they prefere- I read about DMEM + 10 %FCS (or, after plating N2 supplement), but does anyone have experience with the use of RPMI medium + 10% FCS?

DMEM plus 10% (5% also works well) serum is all you need for B104 cells. these cells grow even if you don't want them to grow... don't worry, they are easy to culture. in my experience, only the number of splits (propagations) can lead to a bad growth (do not exceed over 15), anyway, it depends on the individual crio-preservation cell clone conditions.