I got mouse bone-marrow mesenchymal stem cell(passage 1) and fixed and stained them with DAPI and got following images.
This is my first time dealing with mMSC but based on sizes of nucleus I think there are two different cell lines in my culture. and also, i don't know why there are speckles in all nucleus.
Any ideas?
Hello Chi Woo Yoon
I think you should try some later passages may be between (P4-P6), the cells will be much more healthy there and if there is a contamination due to some other population or hematopoietic, it may get rid off.
You may try once.
All the best !!!
Part of what you are seeing may be due to the high density of MSCs in your culture. MSCs partially differentiate in vitro at high densities, especially if the media is not exchanged frequently. I have noted that a cell type with small cell bodies suddenly appears in my rat MSC cultures if I let them go to confluency. I try to passage the MSCs when they get to about 80% confluency. Good luck. Growth rates of individual MSC preparations can be quite variable.
I think Purva is likely right - mouse bone marrow MSCs take until around passage 4 to be pure from hematopoietic cells (bright cells?). I don't know if that is true for rat too, karen? That is unless you cell sorted by surface marker or included MesenPure (from stemcelltech). Incubating at low oxygen is supposed to help them grow if you have the incubator. If you don't (like me) then you will likely have trouble getting them to grow... they go through a transformation (crisis) that also causes chromosomal abnormalities. It takes me around 90 days to get from p1-p4 where they finally proliferate. I understand that this problem is specific to mouse MSCs: human MSCs grow very well without transformation. Karen is also right that high confluency favours differentiation (for me they form adipocytes, which is good because at least it is very easy to see what's happening), but from your images it looks like the large nuclei are your MSCs and they are not at high confluency yet, in my opinion.
How do they look with light microscopy? I attach my unpure MSCs at P1 (mostly hematopoietic, not growing) and P4 (pure, fibroblasts) grown in mesencult in case that helps. They were C57Bl6, every strain is different though.
This papers says about internationalist of synthesized porphyrins and Hoechst staining in candida cells
https://www.researchgate.net/publication/279459630_Axial_Ligand_Modified_High_Valent_Tin%28IV%29_Porphyrins_Synthesis_Structure_Photophysical_Studies_and_Photodynamic_Antimicrobial_Activities_on_Candida_Albicans
Article Axial Ligand Modified High Valent Tin(IV) Porphyrins: Synthe...
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