Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Conditional deletion of Abcb5 in mice (A) Schematic representation of the strategy used to target the Abcb5 gene. LoxP sites (red triangles) were inserted upstream and downstream of exon 2 (E2). The neomycin resistance cassette ( NEO ) is flanked by Frt sites (yellow ovals). (B) After the initial screening of the positive ES cell clones, a 5′ external probe (red rectangle) was used to identify the targeted alleles by detecting a shift from the endogenous HindIII wild-type (WT) band of 13 kb to the rearranged 8.7 Kb band resulting from the insertion of the neo cassette by homologous recombination. The correct targeting was confirmed by using a 3′ external probe following digestion with the HindIII restriction enzyme, which detects a shift from the 13 Kb endogenous WT band to the rearranged 6.6 Kb mutant band. HSV-TK, thymidine kinase cassette for the negative selection; pBS, backbone pbluescript vector. (C) The presence of the distal loxP site in the genome was tested by PCR. Amplification of the WT or mutant Abcb5 alleles resulted in bands of either 239 bp (WT) or 330 bp (recombined allele). (D) Mice were routinely genotyped by isolating genomic DNA from an ear punch using 100 ng of genomic DNA as the template. Amplification of the WT or mutant Abcb5 alleles resulted in bands of either 512 bp (Abcb5 +/+ ) or 112 bp (Abcb5 −/− ).

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Clone Assay, Homologous Recombination, Mutagenesis, Selection, Plasmid Preparation, Amplification

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: The Abcb5-mutant mice have lower lactate levels, indicating a shift in cellular energy metabolism (A) Lactate levels [mmol/L] in female (f) and male (m) Abcb5 +/+ (control, con) and Abcb5 −/− (mutant, mut) mice. Reduced lactate levels were detected in the ABCB5 −/− mice ( p < 0.01), . (B) Fasting cholesterol boxplot with stripchart, split by sex and genotype ( p = 0.046), . (C) Fasting HDL cholesterol boxplot with stripchart, split by sex and genotype ( p = 0.028), . (D) Triglyceride boxplot with stripchart, split by sex and genotype ( p = 0.016), . Two cohorts, each consisting of 10 male and 10 female WT and homozygous mutant adult mice, all born within one week, were subjected to clinical chemistry analyses at 13 and 14 weeks of age. Means, standard deviations, and p -values for genotype, sex, and genotype–sex interaction effects were calculated using a linear model. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Mutagenesis, Control

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Lipid alteration of Abcb5-mutant mice heart and liver (A) The number of heart and liver lipid droplets after staining with Oil Red O ( n = 10). Droplets were counted using ImageJ. Abcb5-deficient mice showed a reduced number of lipid droplets in the heart ( p = 0.011), and a female-specific decrease in droplet number in the liver ( p = 0.021). (B) Diameter of heart and lipid droplets after staining with Oil Red O ( n = 10). The Droplets diameter was determined using ImageJ. A female-specific decrease in droplet size in the liver was observed ( p = 0.019). (C) Representative image of heart sections stained with Oil Red O. (D) Representative image of liver sections stained with Oil Red O. Two cohorts, each consisting of 10 male and 10 female WT and homozygous mutant adult mice, all born within one week, were analyzed for heart and liver lipid content at 8 weeks of age. Means, standard deviations, and p -values for genotype, sex, and genotype–sex interaction effects were calculated using a two-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Scale bar = 100 μM.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Mutagenesis, Staining

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Variation of ketone bodies levels in Abcb5-mutant mice Quantification of ketone bodies (acetoacetate, acetate, β-hydroxybutyrate) in the plasma (A), heart (B), and liver (C). Variations in acetoacetate levels were observed in the liver ( p = 0.028) of Abcb5-knockout mice. A female-specific decrease and a male-specific increase in β-hydroxybutyrate levels was observed in heart ( p = 0.048) and liver ( p = 0.020). A similar observation was made for acetate levels in the liver ( p = 0.028). Two cohorts, each consisting of 10 male and 10 female WT and homozygous mutant adult mice, all born within one week, were analyzed for ketone body levels at 8 weeks of age. Means, standard deviations, and p -values for genotype, sex, and genotype–sex interaction effects were calculated using a two-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See .

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Mutagenesis, Clinical Proteomics, Knock-Out

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: ERα activates ABCB5β transcription MelJuSo were transfected with pSG5-hERα and either 3xERE::luc-TATA, ABCB5β_3xERE::luc-TATA or ABCB5β_0xERE::luc-TATA ( n = 6). 24 h post-transfection, cells were incubated with 5 mM estrogen. Then, luminescence was analyzed using the Dual-Luciferase Reporter Assay System (Promega). The y axis represents the luminescence fold change compared to MelJuSo transfected with pSG5 empty plasmid and 3xERE::luc-TATA in the absence of estrogen.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Transfection, Incubation, Luciferase, Reporter Assay, Plasmid Preparation

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Differential Abc gene expression profile in Abcb5-knockout mice Representation of the impact of Abcb5 KO on the expression of other Abc genes in seven different organs. Points and error bars represent geometric means and 95% confidence intervals on the mean, respectively. Statistical significance is represented using darker dots. The mRNA expression profile of 52 murine Abc transporters was measured in seven tissues collected from 5 male and 5 female WT and homozygous mutant adult mice, all born within one week, and analyzed at 12 weeks of age.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Gene Expression, Knock-Out, Expressing, Mutagenesis

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Heatmap representing the impact of Abcb5 knockout on the expression of other Abc genes in seven different organs Tile colors represent the ΔΔCt values. Low ΔΔCt corresponds to overexpression and is red colored. Genes and organs are clustered following the Euclidean distance; data are not scaled. The mRNA expression profile of 52 murine ABC transporters was measured in seven tissues collected from 5 male and 5 female WT and homozygous mutant adult mice, all born within one week, and analyzed at 12 weeks of age.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Knock-Out, Expressing, Over Expression, Mutagenesis

Journal: Regenerative Therapy

Article Title: Conditioned media of stem cells from human exfoliated deciduous teeth contain factors related to extracellular matrix organization and promotes corneal epithelial wound healing

doi: 10.1016/j.reth.2025.03.002

Figure Lengend Snippet: Immunohistochemical findings of limbal stem cell markers in the limbal epithelium of chronic graft-versus-host disease (cGVHD) mice. Sections of the limbus from syngeneic control mice and cGVHD mice treated with vehicle, stem cells from human exfoliated deciduous teeth-conditioned medium (SHED-CM), immortalized-SHED-CM (IM-SHED-CM), or >3.5 kD fractionated components were stained for ATP-binding cassette sub-family B member 5 (ABCB5) (green) (a) or p63 (green) (b) (n = 5–6, per group). The corneal epithelium is located above the white dotted line. Data are presented as the mean ± standard error of the mean. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. One-way analysis of variance with Tukey–Kramer's post-hoc test was used for the analysis. Scale bar, 10 μm.

Article Snippet: This process was applied to antibodies against p63 (dilution of 1:500, sc-25268, monoclonal, Santa Cruz Biotechnology, Dallas, TX), ATP-binding cassette sub-family B member 5 (ABCB5) (dilution of 1:500, bs-1604R, Bioss Antibodies, Woburn, MA), 4-hydrogy-2-nonenal (4-HNE) (dilution of 1:100, HNEJ-2, Japan Institute for the Control Aging, Shizuoka, Japan), or 8-hydroxy-2′-deoxyguanosine (8-OHdG) (dilution of 1:50, MOG-020P, N45.1, Genox, Torrance, CA).

Techniques: Immunohistochemical staining, Control, Staining, Binding Assay

Journal: Revista Brasileira de Ortopedia

Article Title: Osteopontin, WNT3A, and ABCB5 Biomarkers Expression in Osteosarcoma Patients

doi: 10.1055/s-0044-1788671

Figure Lengend Snippet: Description of primary antibodies, their respective manufacturers, and dilutions

Article Snippet: ABCB5 , Anti-ABCB5, clone 5H3C6 , GeneTex , 1:200.

Techniques:

Journal: Revista Brasileira de Ortopedia

Article Title: Osteopontin, WNT3A, and ABCB5 Biomarkers Expression in Osteosarcoma Patients

doi: 10.1055/s-0044-1788671

Figure Lengend Snippet: Descrição dos anticorpos primários com seus respectivos fabricantes e diluições

Article Snippet: ABCB5 , Anti-ABCB5, clone 5H3C6 , GeneTex , 1:200.

Techniques:

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet: Conditioned medium from ABCB5+ MSCs and ASCs decreases cisplatin-induced apoptosis of ciPTECs. (A) CiPTECs were treated with cisplatin for 1 h and then further 23 h in the absence and presence of MSC conditioned medium (CM). After 24 h, cisplatin-containing medium was replaced with fresh CM or CTRL medium without cisplatin and in the presence of Apotracker. Apoptosis of ciPTECs was monitored by live cell imaging for 3 days. (B) Representative images of ciPTEC day 1, 2 and 3 after cisplatin treatment started. Green: apoptotic cells. Scale bars: 200 µm. (C) Quantification of ciPTEC apoptosis on day 3. The bars indicate mean fold change relative to cisplatin-CTRL; (D) Quantification of free thiol content in MSC-derived CM. One-Way ANOVA with Tukey’s multiple comparisons test; **p<0.01, cisplatin CTRL N=1, ABCB5+ CM N= 3, ASC CM N= 4, ciPTEC n=3.

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Live Cell Imaging, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet: ABCB5+ cells are able to decrease PBMC proliferation in both human and rat. (A) Experimental layout; image created with BioRender. Proliferation index of stimulated (B) hPBMCs and (C) rPBMCs cultured alone or with MSCs. hPBMC cocultures: CTRL N=11, ABCB5+ cells N=5, ASC and rMSC N=11; rPBMC cocultures: CTRL N=11, ABCB5+ cells N=7, ASC N=4, rMSC N=7. Measurement of kynurenine and nitrite in the conditioned media of (D, F) MSCs-hPBMC and (E, G) MSCs-rPBMC cocultures. Kynurenine measurement for hPBMC cocultures: ABCB5+ cells N=7, ASC and rMSC N=8; for rPBMC cocultures: ABCB5+ cells N=5, ASC and rMSC N=4. Nitrite measurement for hPBMC cocultures: ABCB5+ cells N=7, ASC and rMSC N=8; for rPBMC cocultures: ABCB5+ cells N=8, ASC N=4, rMSC N=8. One-Way ANOVA with Tukey’s multiple comparisons test; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Cell Culture

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet: ABCB5+ cells-derived CM shows modest effect on macrophage phagocytosis but suppresses LPS-induced TNF-α secretion in both species. Experiment al layout shown in Figure 4A . (A-D) Phagocytosis capacity of (A) hMac and (B) rMac after CM treatment. hMac N= 4 and rMac N=4, ABCB5+, ASC and rMSC CM N=3. Phagocytosis of hMac (C) and rMac (D) after inhibition of MSC factors in the presence and absence of CM (pooled from N=3 MSC). hMac N=4; rMac N=3. TNF-α secretion of (E) hMac and (F) rMac after 24 h LPS stimulation (100 ng/ml) in the presence and absence of CM. hMac N= 4 and rMac N=4, ABCB5+, ASC and rMSC CM N=3. (G) Simplified diagram depicting M1 and M2 characteristics (Image was created using BioRender). Error bars show maximum and minimum value relative to respective CTRL; One-Way ANOVA (A-D) and Two-Way ANOVA (F, G) with Tukey’s multiple comparisons test; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Derivative Assay, Inhibition

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet: ABCB5+ MSCs do not ameliorate renal injury in rats pretreated with cisplatin. (A) The timeline of ABCB5+ treatment on rats pretreated with cisplatin. (B) Creatinine and (C) urea level in rats’s plasma and (D) ABZWCY-HPβCD half-life on the day before and 2, 7, 14 days after cisplatin treatment with and without ABCB5+ injection. The bars indicate mean value with standard deviation. For pCreatinine and pUrea: healthy CTRL N=3, cisplatin CTRL N=19, cisplatin vehicle CTRL N= 4, cisplatin i.v. ABCB5+ N=12, cisplatin i.p. ABCB5+ N=12; for ABZWCY-HPβCD half-life: healthy CTRL N=3, cisplatin CTRL N=13, cisplatin vehicle CTRL N= 4, cisplatin i.v. ABCB5+ N=11, cisplatin i.p. ABCB5+ N=12. The bars indicate mean value with standard deviation; Mixed-effect analysis; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. (E-H) The morphology of rats’ kidneys on day 15 after cisplatin treatment of the following group: (E) healthy CTRL, (F) cisplatin CTRL, (G) cisplatin i.v. ABCB5+ and (H) cisplatin i.p. ABCB5+. H&E staining. Scale bars: 1 mm. Images acquired with Axio Scan.Z1 microscope (ZEISS). (I–L) Corresponding magnification of the corticomedullary region showing the altered cytoarchitecture due to the induced damage. Light blue arrows: proximal tubular dilatation; blue arrows: dregs of extruded necrotic cells; green arrows: protein casts; yellow arrows: new fibrotic tissue deposition and inflammatory cells infiltrate. H&E staining. Images acquired with Axio Scan.Z1 microscope (ZEISS).

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Clinical Proteomics, Injection, Standard Deviation, Staining, Microscopy

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet: Cisplatin and ABCB5+ cells exert changes in gene expression of rats’ kidney. (A) The gene expression of rats’ kidney on day 15 after cisplatin treatment depicted in a 3D-principal component (PC) analysis graph. (B) The total number of genes that are significantly down- and upregulated in healthy CTRL, cisplatin i.v. and i.p. ABCB5+ groups, compared to cisplatin CTRL group. All groups N=3.

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Gene Expression

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet: ABCB5+ cells promote changes in pathways of cisplatin-treated rats. Gene set enrichment analysis based on KEGG database shows the most upregulated and downregulated pathways of rats treated with cisplatin ABCB5+ cells iv (A) and ip (B) , in comparison with cisplatin CTRL. All groups N=3.

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Comparison

Journal: Frontiers in Immunology

Article Title: Clinical-grade human skin-derived ABCB5+ mesenchymal stromal cells exert anti-apoptotic and anti-inflammatory effects in vitro and modulate mRNA expression in a cisplatin-induced kidney injury murine model

doi: 10.3389/fimmu.2023.1228928

Figure Lengend Snippet:

Article Snippet: Clinical-grade ABCB5+ MSCs were provided by TICEBA GmbH (Heidelberg, Germany) ( ) and were thawed and either seeded directly in the experimental plate the night prior to the experiments or formulated for injection.

Techniques: Control, Modification, Saline, FACS, Filtration, Immunopeptidomics, RNA Sequencing