Murine bone marrow progenitors in cuture - (Nov/05/2009 )

Hi. I'm trying to culture bone marrow cells. My goal is to derive them to a particular celluar type. I've got few protocols about to mainteining them with cytokines but my cells don't live from about day 3 in culture. I want to know if somebody has worked with this progenitors and could give me some advises. I use RPMI medium with BFS 20% and non-essential aminoacids (IX).

Thanks very much

Thanks.

I'm trying to differentiate them into eosinophils. I've found many protocols where they use cytokines like murine SCF and FLT3L since the day 0. They use to IL-5 from day 4 to day 12 in culture. I would like to know if you know how I could maintain them in less day because the yield decreases with the time. So I want to know if you have experience in murine bone marrow cells culturing and you know the conditions to maintain them with the most viablility posible.

Tnahls again!

smyn on Nov 6 2009, 06:38 PM said:

Thanks.

I'm trying to differentiate them into eosinophils. I've found many protocols where they use cytokines like murine SCF and FLT3L since the day 0. They use to IL-5 from day 4 to day 12 in culture. I would like to know if you know how I could maintain them in less day because the yield decreases with the time. So I want to know if you have experience in murine bone marrow cells culturing and you know the conditions to maintain them with the most viablility posible.

Tnahls again!

I have also been attempting something similar to this. I've been following the protocol by Kim Dyer, "Functionally Competent Eosinophils Differentiated Ex Vivo in High Purity from Normal Mouse Bone Marrow". I've had very little sucess to date with this. It's quite frustrating.

Have looked into specialist serum for bone marrow culture - StemCell deal with that kind of stuff. I've tried out their FBS, Horse Serum, H3000 and SFEM that have been formulated and tested for long term myeloid cell culture.

Also have tried various companies and their cytokines.

On another forum someone suggested trying BMGS from Genial Genetics as a supplement to the media to inprove the culture's growth. Haven't tried this yet. Have you had any more sucess with this?!