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. 2025 Jan 28;15:3568. doi: 10.1038/s41598-025-87371-3

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© The Author(s) 2025

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 2 — (A) Comparison of Differentiation Potential of imCMSCs. According to the International Society for Cellular Therapeutics (ISCT) definition of mesenchymal stromal cells (MSCs), the osteogenic, adipogenic, and chondrogenic differentiation abilities of immortalized chorionic mesenchymal stromal cells (imCMSCs) were assessed. The cells underwent differentiation into osteoblasts (×200), adipocytes (×400), and chondroblasts (×200) using alizarin red dye, oil red O dye, and Alcian blue dye, respectively. n = 3. (B) Comparison of karyotypic profiles between the two cell lines. (C) Fluorescence in situ hybridization (FISH) on cells (×400). Red fluorescence is β-actin gene, green fluorescence is the target gene. (D) Soft agar cloning experiments were performed to verify the in vitro safety of imCMSCs. A549 was used as a positive control and 3rd-passage CMSCs as a negative control. Magnification is 100 μm. Red arrows are cells.