Journal: bioRxiv

Article Title: TAR syndrome causal gene RBM8A is critical for embryonic bone development and proper Hedgehog signaling

doi: 10.64898/2026.04.21.718480

Figure Lengend Snippet: A. Immunofluorescence staining of cultured human bone marrow-derived mesenchymal stem cells showing CD90 (green) and RBM8A (red). Merged image (right) demonstrates RBM8A localization in CD90-positive cells. Scale bar = 50 µm. B. Whole-mount immunofluorescence staining of E10.5 mouse embryo section showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is broadly expressed, with high expression in the limb bud. In the neural crest, RBM8A expression is decreased in SOX9-positive regions. Scale bar = 200 µm. C. Whole-mount immunofluorescence staining of E14.5 mouse forelimb autopod sections showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is broadly distributed throughout the autopod, including SOX9-positive cartilage condensation. Scale bar = 200 µm. D. Whole-mount immunofluorescence staining of E14.5 mouse forelimb zeugopod longitudinal tissue section showing RBM8A (red), SOX9 (green), and merged image with DAPI (blue). RBM8A is detected in and around SOX9-positive skeletal condensations, including the central region of the developing cartilage element. Scale bar = 200 µm.

Article Snippet: Human bone marrow-derived MSCs were obtained from ATCC (catalog #: PCS-500-012) and cultured using Bone Marrow-Mesenchymal Stem Cell Basal Medium (ATCC, catalog #: PCS-500-030) with 7% FBS and growth factor supplements (15 ng/ml IGF-1, 125 ng/ml FGF2).

Techniques: Immunofluorescence, Staining, Cell Culture, Derivative Assay, Expressing

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: Morphological changes in BM-MSCs during osteogenic differentiation over 28 days in media with different supplement compositions. Cells were differentiated in DM1–DM4 media. Cell morphology was assessed by bright-field microscopy on days 0, 7, 14, and 28. Representative images show progressive changes in cell shape and density indicative of osteogenic differentiation. Scale bar = 200 μm

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Microscopy

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: Von Kossa staining of BM-MSCs cultured for 28 days in different osteogenic media reveals varying degrees of mineralization. Cells were differentiated in DM1–DM4 media. Black precipitates indicate calcium phosphate deposits, demonstrating the extent of ECM mineralization. Scale bar = 500 μm

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Staining, Cell Culture

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: Alizarin Red staining of BM-MSCs cultured in different osteogenic media (DM1–DM4) at various time points reveals differences in calcium deposition. Cells were cultured for 7, 14, and 28 days in DM1–DM4 media. Red staining indicates the presence of calcium-rich deposits. The most extensive mineralization was observed in DM4, particularly on day 14 and 28, whereas DM2 (lacking DEX after day 7) showed the weakest staining. Scale bar = 200 μm

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Staining, Cell Culture

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: A Quantification of calcium deposition in BM-MSCs cultured in osteogenic media DM1–DM4 using Alizarin Red extraction. Absorbance was measured at 562 nm and expressed in arbitrary units. B ALP activity in BM-MSCs cultured in different osteogenic media (DM1–DM4) at various time points. ALP activity was normalized to total protein content and expressed as enzyme activity per milligram of protein per milliliter of lysate. All data represent means ± standard deviations from three independent biological replicates. Statistical significance was assessed using a two-tailed unpaired t -test and is indicated by asterisks (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Cell Culture, Extraction, Activity Assay, Two Tailed Test

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: Visualization of actin cytoskeleton and mitochondrial network in BM-MSCs undergoing osteogenic differentiation in various media (DM1–DM4). Cells were stained with Phalloidin–TRITC (red) to visualize F-actin filaments and MitoTracker™ Red CMXRos (red) to detect mitochondria. Cell nuclei were counterstained with NucBlue™ (blue). Images were processed using ImageJ software (W. S. Rasband, U.S. National Institutes of Health, Bethesda, MD, USA). Scale bar = 100 μm

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Staining, Software

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: Relative expression levels of osteogenic marker genes during BM-MSCs osteodifferentiation in different supplementation conditions. Transcript levels of A RUNX2 , B ALP , C OCN , and D BMP2 were measured by qRT-PCR at days 7, 14, and 28 in BM-MSCs cultured in osteogenic media DM1–DM4. Gene expression was normalized to the housekeeping gene GAPDH and expressed relative to undifferentiated control levels. All data are presented as means ± standard deviation from three biological replicates. Asterisks indicate statistically significant differences (* P < 0.05, ** P < 0.01; unpaired two-tailed t -test).

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Expressing, Marker, Quantitative RT-PCR, Cell Culture, Gene Expression, Control, Standard Deviation, Two Tailed Test

Journal: BioMedical Engineering OnLine

Article Title: Synergistic effects of dexamethasone and vitamins D and K on mesenchymal stem cell differentiation

doi: 10.1186/s12938-026-01556-z

Figure Lengend Snippet: Overview of experimental workflow. Timeline of BM-MSCs culture, induction of osteogenic differentiation in four different media (DM1–DM4), and time points of sample collection. The diagram illustrates key methodological steps, including morphological assessment, von Kossa and Alizarin Red staining, ALP activity and total protein quantification, immunofluorescence staining (F-actin and mitochondria), and gene expression analysis by qRT-PCR. Sampling was performed at days 0, 7, 14, and 28, as indicated. Created in Biorender.com

Article Snippet: Normal human bone marrow-derived mesenchymal stem cells (BM-MSCs) (ATCC PCS-500-012 TM ) (Sigma Aldrich, St. Louis, MO, USA) were used for experiments.

Techniques: Staining, Activity Assay, Immunofluorescence, Gene Expression, Quantitative RT-PCR, Sampling