PC12-Neurite out growth - less no of cells with neurite (Mar/11/2007 )

all my dear freinds i am facing some problem with my experiment , if any body can help me please try,

i am , incubating my PC12 in conditioned medim of the glial cell , and observing the neurite outgrowth ,

i am getting the neurite but number of cell bearing neurite as well as neurite length is very less , what should i do? for increasing

the the same

Our lab was trying to induce neurite outgrowth in PC-12 cells using NGF. We found that the NGF supplier and more importantly, the Horse Serum supplier we used made a huge difference in ability of these cells to differentiate and form neurites. Have you changed anything in your culture conditions recently?

no i naver changed any thing in my culture condition

Just getting used to work in vitro, I also ask that kind of "silly" questions (there are no "silly" question, except the one not asked) in my lab and in this forum.

After one of those questions, our technician explained me that for astrocyte culturing, she test each new batch of comercial serum. If cells grows well, she orders 80-100 bottles, to keep always the same condition. Because serum composition may vary between each lot with huge effects on the cell cultured.

But I think you Amid are asking for some sort of dendrite formation inducer in PC12 cells, aren't you?

hi,
the answer of vw3sarah22 is very correct!!!
i case you dont change anything in culture conditions you should: conditione the medium from glia in lower volume and/or to collect the medium that was conditioned longer and/or with higher cell number. all these things should elevate the concentration of differentiation-inducing factor in the conditioned medium.
let me know about your progress...