Isolation and characterization of a CD34+ sub-clone in B-cell lymphoma

Abstract

Non-Hodgkin's lymphoma (NHL) is the most common hematological malignancy in the US. Many types remain incurable despite response to initial therapy and achievement of complete remission (CR). Advanced laboratory techniques like multicolor flow cytometry (FCM) and polymerase chain reaction (PCR) have demonstrated persistence of rare malignant cell population post therapy. However, the functional and biological characteristics of this population have not been elucidated. Established B-lymphoma cell lines (B-NHL) and patient-derived samples (PDS) were analyzed using 8-color FCM. CD34 + sub-population was enriched using in vitro exposure to 2-chlorodeoxyadenosine (2-CdA) and by CD34 magnetic beads. Genetic analysis of cell fractions was done by karyotyping and array comparative genomic hybridization (aCGH). Sensitivity to chemotherapy was assayed by short-term in vitro exposure to chemotherapy. Clonogenicity was determined by soft agar colony formation assay, and proliferation was determined using DNA staining with propidium iodide and FCM. FCM demonstrated the presence of a minute sub-clone of monotypic B-cells that express CD34 in B-NHL cell lines (3 of 3) and in PDS (8 of 8). This sub-population enriched up to 50 fold in vitro by exposure to 2-CdA and up to 80% purity by CD34 magnetic bead column isolation. Except for CD34 expression, this population expressed identical phenotype and genotype to parent cells, but was more proliferative, Hoechst 33342-positive, clonogenic, and resistant to chemotherapy compared with the CD34population. The isolated CD34 + monotypic B-cells may contribute to resistance of certain NHL to treatment and should be targeted by potential new drugs for NHL.

Quantitative Spectrofluorometric Assay Detecting Nuclear Condensation and Fragmentation in Intact Cells

“…Hoechst 33342 11,22 , 34580 23 or 33258 21 . Hoechst 33342 has been mostly used for ow cytometry [32][33][34] . On the other hand, H33258 staining has been predominantly used in uorescence microscopy 18,19,35 .…”

Uncovering the Exosomes Diversity: A Window of Opportunity for Tumor Progression Monitoring

“…Additional data indicated that DLBCL-derived exosomes expressed the canonical endosomal markers (Alix, CD81, TSG101, and CD63) and the B-cell marker CD20 [75]. However, conventional diagnosis of B-cell malignancies is based on the up-or down-expression of several biomarkers, such as CD5 [76], CD34 [77], CD123 [78], CD20 [79], and CD138 [80].…”

DETexT: An SNV detection enhancement for low read depth by integrating mutational signatures into TextCNN

“…Most SNV detection methods rely on high sequence read depth and false-positive results appear to increase with the decrease of sequencing depth ( Xiao et al, 2021 ). Although advances in sequencing technologies have reduced the cost of high read depth, the effective depth remains low in the case of prenatal fetal detection ( Alba et al, 2012 ), low purity tumors ( Wilkerson et al, 2014 ), and subclonal structures ( Al-Katib et al, 2020 ). For example, liquid biopsies that identify cancer mutations through blood material have been proposed as a transformative technology for early cancer screening and residual disease monitoring ( Kleftogianniss et al, 2020 ).…”