mouse preosteoblast mc3t3 e1 subclone 4 cells  (ATCC)

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects and mechanisms of iron overload on the proliferation and differentiation of preosteoblastic cells via a 3D microsphere culture system

doi: 10.3389/fbioe.2026.1700858

Figure Lengend Snippet: The 3D microsphere culture system attenuates proliferation and ROS expressions in preosteoblastic cells under iron overload. (A) Live/dead staining of cells cutured on different groups. (B) Intracellular ROS staining of preosteoblastic cells cultured on different groups. (C) The corresponding quantitative analysis of live/dead fluorescence. (D) The corresponding quantitative analysis of fluorescence intensities. (E) Growth curve of preosteoblastic cells cultured on TCP and GM groups with α-MEM without FAC. (F) The corresponding quantitative analysis of ratio of early to late apoptosis. (G) Apoptosis analysis of preosteoblastic cells cultured on different groups via Annexin V-FITC/PI flow cytometry. (H) Cell viability of preosteoblastic cells exposure to 50/300 μM FAC for 24/36/72 h. (I) Cell cycle distribution of preosteoblastic cells cultured on different groups via flow cytometry. (J) The corresponding quantitative analysis of cell cycle phase proportions (G1/G2/S). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Mouse preosteoblastic cells (MC3T3-E1 subclone 4; ATCC) were cultured in α-minimum essential medium (α-MEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Staining, Cell Culture, Fluorescence, Flow Cytometry

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects and mechanisms of iron overload on the proliferation and differentiation of preosteoblastic cells via a 3D microsphere culture system

doi: 10.3389/fbioe.2026.1700858

Figure Lengend Snippet: The 3D microsphere culture system attenuates integrin-mediated mechanotransduction in preosteoblastic cells under iron overload. The immunofluorescence staining of (A) ITGA1 and (B) ITGB1 of cells cultured on different groups. And the corresponding quantitative analysis of the mean fluorescence intensity (MFI) for (C) ITGA1, (D) ITGB1 and (E) F-actin. (F) The mechanotransduction-related gene expressions of cells cultured on different groups. (G) The corresponding quantitative analysis of the absolute reduction of mRNA expression for mechanotransduction-related genes. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mouse preosteoblastic cells (MC3T3-E1 subclone 4; ATCC) were cultured in α-minimum essential medium (α-MEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Immunofluorescence, Staining, Cell Culture, Fluorescence, Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects and mechanisms of iron overload on the proliferation and differentiation of preosteoblastic cells via a 3D microsphere culture system

doi: 10.3389/fbioe.2026.1700858

Figure Lengend Snippet: The 3D microsphere culture system attenuates osteogenic function impairment in preosteoblastic cells under iron overload. The immunofluorescence staining of (A) OPN and (B) OCN of cells cultured on different groups. And the corresponding quantitative analysis of the MFI for (C) OPN, (D) OCN. (E) Alizarin Red (calcium nodule formation), alkaline phosphatase (ALP, early osteogenic marker), and Sirius Red (collagen deposition) staining. Quantification of (F) Alizarin Red staining, (G) ALP staining, (H) Sirius Red. (J–I) The osteogenic - related gene expressions of cells cultured on different groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mouse preosteoblastic cells (MC3T3-E1 subclone 4; ATCC) were cultured in α-minimum essential medium (α-MEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Immunofluorescence, Staining, Cell Culture, Marker

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects and mechanisms of iron overload on the proliferation and differentiation of preosteoblastic cells via a 3D microsphere culture system

doi: 10.3389/fbioe.2026.1700858

Figure Lengend Snippet: RNA sequencing analysis of preosteoblastic cells cultured on GM + FAC and TCP + FAC. (A) Volcano plot of DEGs between GM and TCP groups. (B,C) KEGG pathway enrichment analysis of upregulated and downregulated genes. (D) Cluster heatmap of DEGs in GM and TCP, with low expression indicated in blue and high expression in orange. (E,F) GO enrichment analysis of upregulated and downregulated genes between the two groups. (G–L) GSEA enrichment analysis of inter-group signaling pathways.

Article Snippet: Mouse preosteoblastic cells (MC3T3-E1 subclone 4; ATCC) were cultured in α-minimum essential medium (α-MEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: RNA Sequencing, Cell Culture, Expressing, Protein-Protein interactions

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects and mechanisms of iron overload on the proliferation and differentiation of preosteoblastic cells via a 3D microsphere culture system

doi: 10.3389/fbioe.2026.1700858

Figure Lengend Snippet: RNA sequencing analysis of preosteoblastic cells cultured in the same culture system (TCP vs TCP + FAC, GM vs GM + FAC). (A,B) GO enrichment analysis for upregulated and downregulated genes of TCP + FAC and TCP. (C,D) KEGG pathway enrichment analysis for upregulated and downregulated genes of TCP + FAC and TCP. (E,F) GO enrichment analysis for upregulated and downregulated genes of GM + FAC and GM. (G,H) KEGG pathway enrichment analysis for upregulated and downregulated genes of GM + FAC and GM.

Article Snippet: Mouse preosteoblastic cells (MC3T3-E1 subclone 4; ATCC) were cultured in α-minimum essential medium (α-MEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: RNA Sequencing, Cell Culture

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Effects and mechanisms of iron overload on the proliferation and differentiation of preosteoblastic cells via a 3D microsphere culture system

doi: 10.3389/fbioe.2026.1700858

Figure Lengend Snippet: The schematic diagram of integrin mediated-signal pathway regulations on preosteoblastic cells under iron overload in 3D cell culture.

Article Snippet: Mouse preosteoblastic cells (MC3T3-E1 subclone 4; ATCC) were cultured in α-minimum essential medium (α-MEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Cell Culture