I am culturing hOBs in osteogenic media and post 10-20 days after adding differentiation media morphology of the cells changed and some granular material is deposited around the cells. I would like to know whether these are hydroxyapatite deposits or something else.
I think its waste material , you must wash cells with PBS to remove it
Aqsa Qambrani I used to wash the cells 2-3 times with PBS before changing the media, but it didn't get detached. So, I thought these must be extracellular deposits.
Aqsa Qambrani As, I mentioned these were the deposits that were even not removed by 2-3 times washing with PBS. Moreover for detachment I am using accutase not trypsin.
Hello Khushpreet Kaur ,
have you solved this issue? have you observed that this is a correct osteogenesis or if osteogenic differentiation should look different? because I am currently encountering the very same morphology and so doubt.
I thank you in advance if you will reply,
best,
giulia
Giulia Trimaglio I found these as extracellular deposits. You'll confirm it using Alizarin red staining. Same morphology, I got in differentiating osteoblast cell to mature osteoblasts. Even media gets turbid and arises doubt about contamination. But its not so, these are proteins and matrix deposited by actively proliferating osteoblasts.
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