Recently, I have started facing this issue that when I take some BMDM media just after making it, I see thread like structural cells. So, I tried to filter it using 0.22um and then I started taking it again and plated and those residues or whatever those structures are, are not there anymore. my mice colony has some breeding issue and that is why I can not just use a mice to test if the media is fine or not. It would be awesome if you guys could help me out :
1. Am i really dealing with contamination ?
2. If yes, is filtration enough to get rid of the contamination ? because I did not see those after filtering.
3. If I find out later that it is contamination, how is it possible that every time I am making a new bottle using all new aliquot, it is still there ?
4. If it is because of the aliquot of L929 cell conditioned media, can I use something else to replace L929 conditioned media ?
Please help !
Hi Antara Roy . Sorry, I have seen many questions on RG regarding BMDM through the years, most with few answers. Maybe contact the researchers directly with your dilemma.
https://www.researchgate.net/post/Culture_and_Proliferation_of_BMDM_Bone_Marrow_Derived_Macrophages
https://www.researchgate.net/post/Hi_i_have_a_question_about_BMDM_please_After_6_days_differentiate_should_I_use_media_containing_MCSF_to_culture_BMDM_for_the_following_experiment
https://www.researchgate.net/post/Any-experience-with-passaging-of-murine-BMDM-Do-you-keep-L929-in-your-medium-after-passaging-cells
Dear Antara,
difficult to say, if you really have a contamination or if you just see some fibroblasts, which always come by with the preparation.
If you like you can have a look at our very detailed protocol for preparation, differentiation and cultivation of BMDM:
Article Highly Efficient Transfection of Primary Macrophages with In...
I hope that helps. Please ask any time, if you have any further questions.
All the best and stay healthy,
Marc
alright, i will take a look into everything that you guys have suggested. Thank you Marc Herb Steingrimur Stefansson
Hi, do you have any suggestions on the seeding density ratio for bone marrow derived macrophages extracted from rabbit?