Hi there fellow respected researchers and industry players. I am currently facing a problem with isolating mesenchymal stem cells from menstrual blood. To address the concern of contamination from the natural flora, my protocol included a washing step to wash the blood sample using a PBS solution supplemented with antibiotics and antifungals. This step was repeated for four times before proceeding to Ficoll-paque isolation. Risk of contamination was greatly under control but I observed that the number of mesenchymal cells isolated were very low. My question is, is anyone doing menstrual blood mesenchymal cell also experiencing low yield from isolation? Any suggestions as to how I may improve the yield of menstrual blood isolated mesenchymal cells is greatly appreciated.
I would directly add the menstrual blood sample to culture flasks with medium containing antibiotics and antifungal agents to determine a baseline for colony forming cells. Change medium on day 3 and count colonies on day 6-7. After that you can determine how many potential MSCs are in your sample and whether contamination is a problem. If so, start with one wash or start with a density gradient that will also remove many contaminants.
Thank you Dr Pittinger for your suggestion. I shall try it out. I have not tried directly culturing the sample, it'd be great if this works!
Have a nice day.
I had this problem for bone derived MSCs. I increased duration of experiment and exchange of 2/3 medium. MSCs is producing growth factores, so dont throw all. The problem was solved. Sincerely.
Kemal Yuce per your recommendation, I am seeing some improvement in the isolation and growth. I still have some improvements to do as I feel it is still not very robust. If there is any more tips, would greatly appreciate it! thank you for your recommendation! one more question, you mentioned that you increased duration of experiment, could you elaborate more? perhaps, if information is sensitive, we can communicate via email
Have a nice day.
my e mail is [email protected]. With regard to your quations, if ı have information, ı can aswer.
with kind regards.
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