magnetic beads conjugated to cd138 antibodies  (Miltenyi Biotec)

Journal: International Journal of Oncology

Article Title: LGR4 promotes proliferation and homing via activation of the NF-κB signaling pathway in multiple myeloma

doi: 10.3892/ijo.2025.5718

Figure Lengend Snippet: High expression of LGR4 is associated with cell adhesion and poor prognosis in multiple myeloma. (A) Gene expression heatmap of LGR4 (Red) and Wnt/β-catenin signal related genes (Black), Cell adhesion associated genes (Blue) in CD138 + cells from HD (n=22), MGUS (n=44) and MM (n=351). (B) Pearson's correlation analysis of the relationship between LGR4 and cell adhesion genes. Red for positive, blue for negative. (C) Gene expression heatmap of OCI-My5 and OCI-T3 rd -luc with LGR4 (Red), Wnt/β-catenin signal genes (Black), Cell adhesion associated genes (Blue). (D) Gene expression heatmap of KMS28-BM and KMS28-PE with LGR4 (Red), Wnt/β-catenin signal genes (Black), Cell adhesion associated genes (Blue). (E) Representative images of immunofluorescence images of LGR4 and ZEB1 protein expression in KMS28-BM and KMS28-PE. Scale bars, 50 μ m. (F) Kaplan-Meier analyses of overall survival in MM patients with LGR4 low ZEB1 low (n=58), LGR4 low ZEB1 high (n=7), LGR4 high ZEB1 low (n=309) and LGR4 high ZEB1 high (n=85) from GSE2658. HD, healthy donors; MGUS, monoclonal gammopathy of undetermined significance; MM, multiple myeloma; BM, bone marrow; PE, pleural effusion; ZEB1, Zinc Finger E-Box Binding Homeobox 1.

Article Snippet: CD138 + plasma cells were isolated by using anti-human CD138 magnetic beads (Miltenyi Biotec GmbH) and incubated in 4°C for 15 min with monocytes isolated from BM samples using lymphocyte separation medium (cat. no. LTS1077; TBD; https://www.tbdscience.com/ ).

Techniques: Expressing, Gene Expression, Immunofluorescence, Binding Assay

Journal: International Journal of Oncology

Article Title: LGR4 promotes proliferation and homing via activation of the NF-κB signaling pathway in multiple myeloma

doi: 10.3892/ijo.2025.5718

Figure Lengend Snippet: Overexpression of LGR4 promotes cells' homing to the BM and MM progression in vivo . (A) Schematic of in vivo experiments. (B) Tumor-associated live imaging of NCG mice injected with OCI-Ctrl or OCI-LGR4-OE cells at 4 and 6 weeks (n=5 for each group). (C) Quantification of luminescence intensity in live NCG mice. (D) Flow cytometric analysis images and statistics of the human MM cell proportion in the bone marrow after sacrificing NCG mice. (E) Micro-CT images of tibia derived from NCG mice. (F) Quantification of bone microstructural parameters, namely BV/TV, Tb.N, Tb.Th and Tb.Sp (n=3). (G) TRAP staining for NCG xenografted mice bone marrow section. Scale bars, 200, 25 and 10 μ m. (I) The quantification of the number of Trap-positive osteoclast cells is presented in the column graph. (H) Neoplastic CD138-positive plasma cells. Scale bars, 10 μ m. (J) The quantification of the number of neoplastic CD138 positive plasma cells is presented in the column graph. Statistical analyses were performed using Student's t-test. * P<0.05, ** P<0.01, *** P<0.001 and ****P<0.0001. BM, bone marrow; MM, multiple myeloma; LGR4-OE, LGR4 overexpression; BV/TV, trabecular bone volume fraction; Tb.N, trabecular number; Tb.Th, trabecular thickness; Tb.Sp, trabecular separation; TRAP, tartrate-resistant acid phosphatase; EV, empty vector; ns, not significant (P>0.05).

Article Snippet: CD138 + plasma cells were isolated by using anti-human CD138 magnetic beads (Miltenyi Biotec GmbH) and incubated in 4°C for 15 min with monocytes isolated from BM samples using lymphocyte separation medium (cat. no. LTS1077; TBD; https://www.tbdscience.com/ ).

Techniques: Over Expression, In Vivo, Imaging, Injection, Micro-CT, Derivative Assay, Staining, Clinical Proteomics, Plasmid Preparation

Journal: Biosensors

Article Title: Dual Biomarker Strategies for Liquid Biopsy: Integrating Circulating Tumor Cells and Circulating Tumor DNA for Enhanced Tumor Monitoring

doi: 10.3390/bios15020074

Figure Lengend Snippet: Combined analysis of circulating tumor cells and circulating tumor DNA for the diagnosis and molecular profiling of melanoma, urothelial cancer, and pancreatic cancer.

Article Snippet: MM , 139 MM patients (107 cfDNA, 56 CTC) , Anti-CD138 magnetic bead-based positive selection , Clonal somatic mutations and CNAs detected using ULP-WGS and WES , Qiagen Circulating Nucleic Acids Kit , Clonal somatic mutations and CNAs detected using ULP-WGS and WES , CTCs and cfDNA demonstrated complementary profiles, with some mutations unique to each. Sequential cfDNA monitoring correlated with disease progression and therapeutic responses. , [ ] .

Techniques: Biomarker Discovery, Isolation, Extraction, Selection, Staining, DNA Extraction

Journal: Nature Communications

Article Title: Deeper response predicts better outcomes in high-risk-smoldering-myeloma: results of the I-PRISM phase II clinical trial

doi: 10.1038/s41467-024-55308-5

Figure Lengend Snippet: A Time to progression or last follow-up for patients with sequencing data (one patient per row, grouped by progression status). Green boxes indicate baseline sequencing data available for each patient. Vertical dotted line denotes end of treatment. B Uniform manifold approximation and projection (UMAP) of patient and HD plasma cells that passed quality filtering. Colors indicate individual patients or HD as a group C UMAP colored by malignancy status (tumor vs normal cells). D UMAP colored by biochemical progression status. E Volcano plot of differential gene expression in tumor cells of progressors ( n = 10) and non-progressors ( n = 3). Two-sided p values were computed with Wilcoxon rank-sum test and corrected using the Benjamini-Hochberg approach. Stars = top 30 genes on either side of the volcano with q < 0.05. F Mean HLA class I gene expression from tumor bulk RNA-seq from the PADIMAC cohort who responded (Bortezomib_good, n = 13) or did not respond (Bortezomib_standard, n = 31) to Bortezomib treatment. Box: 1st quartile, median, 3rd quartile; whiskers: ±1.5*interquartile range. The p value was computed with two-sided Wilcoxon rank-sum test. G UMAP of T cell subtypes ( H ) Average proportion of clonotypes belonging to four clone size categories ( rare , < 1%; small, ≥ 1% and < 5%; medium, ≥ 5% and < 10%; large: ≥ 10%) per patient in baseline BM CD138- samples. Error bars = SD from 100 iterations. P values were obtained using two-sided bootstrapping test to compare the mean proportion of rare clones between progressors and non-progressors. In all types of statistical analysis values of p < 0.05 were considered significant (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). I GZMB+ CD8+ T effector memory cell abundance in expanded clones from non-progressors and progressors. Plot statistics same as ( F ). J UMAP of baseline BM T cells from progressors and non-progressors with matched TCR data. T cells with expanded clones (frequency > 1%) are colored by cell type. Gray = cells with rare clonotypes K Volcano plot of differential gene expression of clonally expanded CD8+ TEM in progressors and non-progressors. Statistics same as ( E ). HD healthy donors, P patient, WGS whole-genome sequencing, HRD hyperdiploidy. Source data are provided as a file.

Article Snippet: Plasma cells were enriched from the bone marrow samples using CD138 magnetic bead separation using single-column positive selection (Miltenyi Biotec, Cat #130-097-614).

Techniques: Sequencing, Clinical Proteomics, Gene Expression, RNA Sequencing, Clone Assay