Hello everybody. I am trying to differentiate SaOS2 (osteosarcoma cell line) into human osteoblast and osteocytes (booth). Does anyone know any specific marker for this cell types. How can I be really sure that after the differentiation process the functionality of the cells also change and not only the morphology?
Another Problem, when I seed my cells for the differentiation. First I seed them with Alpha mem + 10% FBS +10mM HEPES medium. Once they are confluent I add to this medium beta glycerol and L-ascorbic acid. It is supposed that the differentiation process should start here, but my cells continue proliferating and they start to detach and overgrowing, this makes difficult to work with.
Does anyone have any Suggestion for this? I would like to work with human osteoblast and osteocyte cell lines.
Hi Marina,
1. please go thru SaOS2 Osteosarcoma cells as an in vitro model for studying the transition of human osteoblasts to osteocytes (Prideaux M et al)
2. MARKERS:
for OSTEOBLASTS>>
Osteocalcin (OC), core binding factor 1 (CBFA1), collagen type 1 (Coll1), alkaline phosphatase (ALP), nucleostemin (NS), and surface proteins CD 10, CD44, CD 59 and CD 105 can be analyzed using RT-PCR, immunofluorescence and Western blot.
for OSTEOCYTES>>
-Elevated E11: an early osteocyte marker.
-Sclerostin: Expressed by more mature osteocytes
Best,
S
Hello
Added to Swarnendra Singh answer , Look at this article.
Good Luck
https://www.ncbi.nlm.nih.gov/pubmed/24916279
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