Journal: Journal of Cellular and Molecular Medicine

Article Title: Bone marrow mesenchymal stem cells enhance autophagy and help protect cells under hypoxic and retinal detachment conditions

doi: 10.1111/jcmm.15008

Figure Lengend Snippet: Coculturing with BMSCs promotes better cell morphology and fewer apoptotic cells. (A) Culturing in hypoxic conditions reduced the cell viability of 661w cells, which could be rescued by coculturing with BMSCs, although slightly lower than that of normal cells. (B) Red fluorescence indicated JC‐1 in normal cells with a normal Δ Ψm, while green fluorescence indicated a low Δ Ψm in hypoxic cells. Magnification: 20×. (C) Coculture with BMSCs decreased the apoptosis and necrosis in 661w cells undergoing hypoxia. These assays were repeated for three times. *: P < .05, compared with hypoxic cells. #: P < .05, compared with normal cells

Article Snippet: Mouse BMSCs (P6) were bought from Cyagen BioTech, and P7 and P8 cells were used for coculture assays.

Techniques: Fluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Bone marrow mesenchymal stem cells enhance autophagy and help protect cells under hypoxic and retinal detachment conditions

doi: 10.1111/jcmm.15008

Figure Lengend Snippet: Coculturing with BMSCs increased autophagy in 661w cells, and the effect of BMSCs was weakened when autophagy was inhibited. (A) MDC staining showed higher number of autophagosome in hypoxic cells, as compared to normal cells, and the autophagosome was further increased in the cocultured group. Magnification: 20×. (B) In the cocultured group, the expression of LC3II and the ratio of LC3II/LC3I were obviously increased, while the expression of p62 was decreased, compared with those in the hypoxia and normal groups. (C) 3‐MA, an autophagy inhibitor strongly diminished the protective effects of BMSCs. In the autophagy‐inhibited coculture (inhibited) group, the cell viability was decreased to nearly that in the hypoxia group. These assays were repeated for three times. *: P < .05, compared with the other two groups. #: P < .05, compared with cocultured cells

Article Snippet: Mouse BMSCs (P6) were bought from Cyagen BioTech, and P7 and P8 cells were used for coculture assays.

Techniques: Staining, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Bone marrow mesenchymal stem cells enhance autophagy and help protect cells under hypoxic and retinal detachment conditions

doi: 10.1111/jcmm.15008

Figure Lengend Snippet: BMSC transplantation attenuated apoptosis and activated autophagy in photoreceptors in retinal detachment. (A) TUNEL staining showed that the subretinal transplantation of BMSCs significantly reduced the number of TUNEL‐positive cells in the ONL at 3 (cells/mm 2 , P < .05) and 7 d (cells/mm 2 , P < .05) after treatment compared to those in the blank and NC groups. Magnification: 40×. (B and C) At 3 d after transplantation, the treated group showed significantly lower cleaved caspase‐3 levels than the other two groups ( P < .05), and at 1 wk after transplantation, this level decreased to nearly that in the normal retina. Significant differences in caspase‐8 and cleaved caspase‐9 levels were also found between retinas treated with BMSCs and the other two groups. (D) The expression of the microtubule‐associated protein LC3‐II was decreased, and p62 expression was increased at 6 and 10 d after RD compared with those at 3 d after RD, suggesting that autophagy levels peaks on day 3 after RD. Three days after treatment, significantly increased LC3‐II expression and decreased p62 levels were found in the BMSC‐treated group compared to those in the other two groups. However, after 7 d post‐treatment, the levels did not differ significantly among the groups. These assays were repeated for three times

Article Snippet: Mouse BMSCs (P6) were bought from Cyagen BioTech, and P7 and P8 cells were used for coculture assays.

Techniques: Transplantation Assay, TUNEL Assay, Staining, Expressing

Journal: Frontiers in Immunology

Article Title: The role of WTAP in regulating macrophage-mediated osteoimmune responses and tissue regeneration in periodontitis

doi: 10.3389/fimmu.2024.1423378

Figure Lengend Snippet: Reducing of WTAP expression in macrophages promotes BMSC osteogenic differentiation. (A) Schematic diagram of the indirect co-culture system between BMDMs and BMSCs. (B-D) RT-PCR analysis quantifying the mRNA levels of osteogenic markers runx2 , bmp2 , ocn , opn and opg in BMSCs co-cultured with conditional medium at 4 (B) , 7 (C) and 14 (D) days (n = 3). (E) Representative images of ALP staining of BMSCs cultured in conditional medium for 14 days. Scale bar = 500 μm. (F) Quantitative analysis of ALP activity of BMSCs cultured in conditional medium for 14 days (n = 3). (G) Representative images of ARS staining of BMSCs cultured in conditional medium for 21 days. Scale bar = 500 μm. (H) Quantitative analysis of ARS staining in BMSCs (n = 3). Data are presented as the mean ± SD from at least three independent experiments. P values were calculated using two-tailed Student’s t test; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Macrophage-conditioned medium was prepared using the supernatants from shNC- and shWTAP-treated BMDMs, and mixed with osteogenic induction medium for mouse BMSCs (MUXMX-90021, Cyagen) at a ratio of 1:2.

Techniques: Expressing, Co-Culture Assay, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Staining, Activity Assay, Two Tailed Test

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Trans-portal hepatic infusion of cultured bone marrow-derived mesenchymal stem cells in a steatohepatitis murine model

doi: 10.3164/jcbn.20-88

Figure Lengend Snippet: Quantification of liver fibrosis. Luciferase positive bone marrow-derived mesenchymal stem cells (Luc-BMSCs) adjusted to 1.0 × 10 6 cells/100 µl were infused into the spleen of a modified steatohepatitis murine model at 12 weeks of the choline-deficient, l -amino acid-defined diet (12W), and at 4 weeks after infusion, the animals were sacrificed ( n = 8). Liver fibrosis and liver steatosis were similarly induced in a control group that was infused with phosphate-buffered saline (PBS) into the spleen, and after 4 weeks, the animals were sacrificed ( n = 8). The evaluation of liver fibrosis is shown. (A) The mean level of Sirius red-stained area was significantly lower in the BMSC group. Control group vs BMSC group = 5.4 ± 0.9% vs 1.5 ± 0.2%. Error bars indicate SD, n = 8 mice each. ** p <0.01. Magnification ×40, scale bars = 100 µm. (B) Western blotting of α-smooth muscle actin (α-SMA) protein in the liver between the BMSC and control groups. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was used as an internal control, n = 3 each. (C) mRNA expression of collagen1a1 ( Col1a1 ) normalized with β-actin ( Actb ), n = 8 mice each. ** p <0.01.

Article Snippet: B6 mouse-derived BMSCs (passage 6) purchased from Cyagen Biosciences Inc. (Tokyo, Japan) were transduced with lentiviral vectors (#79692-G; BPS Bioscience, San Diego, CA) expressing luciferase, and passage 8 BMSCs (Luc-BMSCs) were used in the experimentation.

Techniques: Luciferase, Derivative Assay, Modification, Control, Saline, Staining, Western Blot, Expressing

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Trans-portal hepatic infusion of cultured bone marrow-derived mesenchymal stem cells in a steatohepatitis murine model

doi: 10.3164/jcbn.20-88

Figure Lengend Snippet: Quantification of liver steatosis and lipid metabolism in the liver. Luciferase positive bone marrow-derived mesenchymal stem cells (Luc-BMSCs) adjusted to 1.0 × 10 6 cells/100 µl were infused into the spleen of a modified steatohepatitis murine model at 12 weeks of the choline-deficient, l -amino acid-defined diet (12W), and, at 4 weeks after infusion, the animals were sacrificed ( n = 8). (A) Hematoxylin and eosin (HE) staining. The lipid droplet deposition area was significantly lower in the BMSC group (control group vs BMSC group = 34.7 ± 2.1% vs 21.2 ± 3.3%). Error bars indicate SD, n = 8 mice each. ** p <0.01. Magnifications ×40 and ×100. (B) mRNA expressions of genes related to lipid metabolism in the liver. n = 8 mice each. * p <0.05, ** p <0.01, ns, not significant. Error bars indicate SD.

Article Snippet: B6 mouse-derived BMSCs (passage 6) purchased from Cyagen Biosciences Inc. (Tokyo, Japan) were transduced with lentiviral vectors (#79692-G; BPS Bioscience, San Diego, CA) expressing luciferase, and passage 8 BMSCs (Luc-BMSCs) were used in the experimentation.

Techniques: Luciferase, Derivative Assay, Modification, Staining, Control

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Trans-portal hepatic infusion of cultured bone marrow-derived mesenchymal stem cells in a steatohepatitis murine model

doi: 10.3164/jcbn.20-88

Figure Lengend Snippet: Assessment of the effects of cultured bone marrow-derived mesenchymal stem cells (BMSCs) on steatosis in HepG2 cells. (A) Oil-red O staining of HepG2 cells (magnification ×40, ×100). Control group: HepG2 cells were exposed to 0.5 mM free fatty acid (FFA) mixture for 72 h in the 12-well plate. BMSC co-culture group: HepG2 cells were exposed to 0.5 mM FFA mixture for 24 h. Then, 5 × 10 4 BMSCs were seeded onto the 0.4-µm-pore size Cell Culture Insert and placed into the 12-well plate with the HepG2 cells, and these two kinds of cells were cultured for another 48 h. Scale bars = 100 µm. (B) Absorbance (492 nm) of Oil-red O staining solution. The mean absorbance at 492 nm of Oil-red O staining solution extracted from wells is shown ( n = 12 per group). Control group vs BMSC co-culture group = 1.4 ± 0.1 vs 1.2 ± 0.1. Error bars indicate SD, n = 12 each, ** p <0.01.

Article Snippet: B6 mouse-derived BMSCs (passage 6) purchased from Cyagen Biosciences Inc. (Tokyo, Japan) were transduced with lentiviral vectors (#79692-G; BPS Bioscience, San Diego, CA) expressing luciferase, and passage 8 BMSCs (Luc-BMSCs) were used in the experimentation.

Techniques: Cell Culture, Derivative Assay, Staining, Control, Co-Culture Assay, Pore Size