Huang, Chung-YouChung-YouHuangLin, Win-LiWin-LiLin2025-11-042025-11-042025-12https://scholars.lib.ntu.edu.tw/handle/123456789/733375Background: Osteogenic differentiation refers to the process by which mesenchymal stem cells (MSCs) transform into bone tissues. The microplate reader, characterized by its requirement for fewer samples, simple operation, excellent repeatability, and rapid detection speed, has been utilized to measure the absorbance of cells undergoing osteogenic differentiation. Research design and methods: In this study, we employed the microplate reader to analyze the histological stains on Wharton’s Jelly Mesenchymal Stem Cells (WJMSCs) in vitro, aiming to streamline future osteogenic differentiation studies. Results: It was not recommended to use a microplate reader to read Optical Density (OD) values for Alizarin red S and Picro-Sirius red (PSR) staining. However, a microplate reader could be effectively used to read OD values at wavelengths of 596, 620, and 680 nm from day 3 to day 5 in von Kossa staining (VKS) results. Conclusions: Thus, while the microplate reader is recommended for early-stage VKS quantification, its use is not advised for Alizarin red S staining (ARS) or PSR assays.enAlizarin red SPicro-Sirius redWharton’s Jelly mesenchymal stem cellsmicroplate readeroptical density valuesosteogenic differentiationvon Kossa staining[SDGs]SDG3journal article10.1080/20565623.2025.256716941031803