I would like to know which in vitro assays are considered most reliable and commonly used to assess osteogenic potential, such as:
- Cell adhesion and proliferation
- Alkaline phosphatase (ALP) activity
- Mineralization (e.g., Alizarin Red S staining)
- Expression of osteogenic markers (e.g., RUNX2, OCN, OPN)
Are there specific cell lines you would recommend (e.g., pre-osteoblasts, mesenchymal stem cells), and what incubation timeframes are typically used for such studies?
Any insights, protocols, or references to key studies would be greatly appreciated.
Thank you in advance!
Best regards, khizra aleena MS Physics Student | [ University of sialkot]
Hi!
MSCs from Lonza work the best you can also look for other brands though if you don't want to do in house isolation.
For MSC differentiation protocol you can either buy osteogenic differentiation medium from Stem Cell Technologies and follow their protocol. If you want to do differentiation yourself here's what one of my ex-lab members performed-
hMSCs were differentiated into osteogenic lineage on tissue culture plate using osteogenic differentiation medium formulated in α-MEM with 10% FBS, 1% penicillin–streptomycin, 10 mM Sodium β-glycerophosphate, 50 μM L-ascorbic acid and 0.1 μM dexamethasone. Osteogenic differentiation protocol was started with 60-70% confluent hMSCs (20,000 cells in 24 well plate) by replacing complete growth medium with osteogenic differentiation medium. For first three days, half of the differentiation medium was replenished daily and on alternate days afterwards till day 15.
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