Massive reorganization of the genome during primary monocyte differentiation into macrophage

2020 ◽  
Vol 52 (5) ◽  
pp. 546-553
Author(s):  
Zhipeng Zhang ◽  
Qi Wang ◽  
Yulong Liu ◽  
Qiu Sun ◽  
Hua Li ◽  
...  
Keyword(s):  
Structural Changes ◽  
Cultured Cells ◽  
Genomic Structure ◽  
Leukemia Cell ◽  
Immunological Response ◽  
Leukemia Cell Line ◽  
Primary Cells ◽  
Chromosomal Structure ◽  
Monocyte Differentiation ◽  
Primary Monocyte

Abstract Monocyte-to-macrophage trans-differentiation has long been studied to better understand this immunological response and aspects of developmental processes more generally. A key question is the nature of the corresponding changes in chromatin conformation and its relationship to the transcriptome during this process. This question is especially intriguing since this trans-differentiation is not associated with progression through mitosis, often considered a necessary step for gross changes in chromosomal structure. Here, we characterized the transcriptional and genomic structural changes during macrophage development of primary human monocytes using RNA-seq and in situ Hi-C. We found that, during this transition, the genome architecture undergoes a massive remodeling to a degree not observed before between structured genomes, with changes in ~90% of the topologically associating domains (TADs). These changes in the TADs are associated with changed expression of immunological genes. These structural changes, however, differ extensively from those described recently in a study of the leukemia cell line, THP-1. Furthermore, up-regulation of the AP-1 family of genes that effected functionally important changes in the genomic structure during the differentiation of the THP-1 cells was not corroborated with the primary cells. Taken together, our results provide a comprehensive characterization of the changes in genomic structure during the monocyte-to-macrophage transition, establish a framework for the elucidation of processes underlying differentiation without proliferation, and demonstrate the importance of verifying with primary cells the mechanisms discovered with cultured cells.

scholarly journals Monocytic THP-1 Cells Diverge Significantly From Their Native Counterparts: A Comparative Examination of The Chromosomal Conformations And Transcriptomes

2021 ◽  
Author(s):  
Yulong Liu ◽  
Hua Li ◽  
Daniel M Czajkowsky ◽  
Zhifeng Shao
Keyword(s):  
Cell Lines ◽  
Chromatin Structure ◽  
Immune Cell ◽  
Leukemia Cell ◽  
Model Systems ◽  
Leukemia Cell Line ◽  
Primary Cells ◽  
Monocytic Leukemia ◽  
Altered Expression ◽  
Wide Range

Abstract Immortalized cell lines have long been used as model systems to systematically investigate biological processes under controlled and reproducible conditions, providing insights that have greatly advanced cellular biology and medical sciences. Recently, the widely used monocytic leukemia cell line, THP-1, was comprehensively examined to understand mechanistic relationships between the 3D chromatin structure and transcription during the trans-differentiation of monocytes to macrophages. To corroborate these observations in primary cells, we analyze in situ Hi-C and RNA-seq data of human primary monocytes and their differentiated macrophages in comparison to that obtained from the monocytic/macrophagic THP-1 cells. Surprisingly, we find significant differences between the primary cells and the THP-1 cells at all levels of chromatin structure, from loops to topologically associated domains to compartments. Importantly, the compartment-level differences correlate significantly with transcription: those genes that are in A-compartments in the primary cells but are in B-compartments in the THP-1 cells exhibit a higher level of expression in the primary cells than in the THP-1 cells, and vice versa. Overall, the genes in these different compartments are enriched for a wide range of pathways, and, at least in the case of the monocytic cells, their altered expression in certain pathways in the THP-1 cells argues for a less immune cell-like phenotype, suggesting that immortalization or prolonged culturing of THP-1 caused a divergence of these cells from their native counterparts. It is thus essential to reexamine phenotypic details observed in cell lines with their native counterparts so as to ensure a proper understanding of functional cell states in vivo.

scholarly journals Monocyte nonspecific esterase: purification and subunit structure

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 479-487 ◽  
Author(s):  
J Yourno
Keyword(s):  
Gel Electrophoresis ◽  
Cultured Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Nonspecific Esterase ◽  
Subunit Structure ◽  
Protein Chain ◽  
Cell Extracts

Abstract Monocyte nonspecific esterase has been purified from cultured cells of the acute myeloid leukemia cell line, ML-1. The purified enzyme shows the characteristic properties of the monocyte neutral serine carboxyl esterase, with high sensitivity to organophosphorus inhibitors and sodium fluoride inhibitor. The enzyme is a membrane protein which in the native state exists as a monomer of a mol wt of approximately 68,000 and a trimer of mol wt 205,000. These forms exhibit a complex pattern of dissociation and reassociation based on apparent noncovalent binding of subunits. The delipidated dissociated enzyme runs as a single protein chain of a mol wt of approximately 62,000 on sodium dodecyl sulfate (SDS) gel electrophoresis. The relation of the subunits to monocyte isoenzymes seen on isoelectric focusing (IEF) and polyacrylamide gel electrophoresis at pH 9.5 (pH 9.5 PAGE) of cell extracts is demonstrated. Availability of purified enzyme allows development of monoclonal antibodies and analysis of myeloid differentiation. In addition, the substrate specificity and function of the purified monocyte ectoenzyme are being examined.

scholarly journals Establishment and characterization of a new human eosinophilic leukemia cell line

Blood ◽  
1985 ◽  
Vol 66 (6) ◽  
pp. 1233-1240 ◽  
Author(s):  
H Saito ◽  
A Bourinbaiar ◽  
M Ginsburg ◽  
K Minato ◽  
E Ceresi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Philadelphia Chromosome ◽  
Leukemia Cell ◽  
Culture Conditions ◽  
Membrane Receptors ◽  
Leukemia Cell Line ◽  
Standard Culture

Abstract A human eosinophilic leukemia cell line, designated as EoL, was established from the peripheral blood of a patient with Philadelphia chromosome-negative eosinophilic leukemia (EL). The EoL cell line grows in single cell suspension with a doubling time of 48 hours for about one year. The reactivity of these cells was tested with a panel of monoclonal antibodies; they were found to express surface IA antigen, myeloid antigen (IF10, MY9) and membrane receptors for interleukin 2 (IL-2, Tac antigen). Under standard culture conditions, a small percentage of cells having more typical eosinophilic characteristics was present. These cells had cytoplasmic granules and were positive for Luxol-fast-blue and eosinophil peroxidase. Under culture conditions to induce the maturation of myeloid cells, such as alkaline medium or addition of dimethyl sulfoxide (DMSO), the frequency of cells with typical eosinophilic features increased to about 40%. In addition, cytogenetic studies showed that cultured cells and original leukemic blasts presented similar chromosome abnormalities. EoL seems to be a unique leukemic line committed to the eosinophilic lineage and can provide a useful in vitro model for the study of malignant eosinophilic properties.

scholarly journals Establishment and characterization of a new human eosinophilic leukemia cell line

Blood ◽  
1985 ◽  
Vol 66 (6) ◽  
pp. 1233-1240 ◽  
Author(s):  
H Saito ◽  
A Bourinbaiar ◽  
M Ginsburg ◽  
K Minato ◽  
E Ceresi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cultured Cells ◽  
Interleukin 2 ◽  
Philadelphia Chromosome ◽  
Leukemia Cell ◽  
Culture Conditions ◽  
Membrane Receptors ◽  
Leukemia Cell Line ◽  
Standard Culture

A human eosinophilic leukemia cell line, designated as EoL, was established from the peripheral blood of a patient with Philadelphia chromosome-negative eosinophilic leukemia (EL). The EoL cell line grows in single cell suspension with a doubling time of 48 hours for about one year. The reactivity of these cells was tested with a panel of monoclonal antibodies; they were found to express surface IA antigen, myeloid antigen (IF10, MY9) and membrane receptors for interleukin 2 (IL-2, Tac antigen). Under standard culture conditions, a small percentage of cells having more typical eosinophilic characteristics was present. These cells had cytoplasmic granules and were positive for Luxol-fast-blue and eosinophil peroxidase. Under culture conditions to induce the maturation of myeloid cells, such as alkaline medium or addition of dimethyl sulfoxide (DMSO), the frequency of cells with typical eosinophilic features increased to about 40%. In addition, cytogenetic studies showed that cultured cells and original leukemic blasts presented similar chromosome abnormalities. EoL seems to be a unique leukemic line committed to the eosinophilic lineage and can provide a useful in vitro model for the study of malignant eosinophilic properties.

scholarly journals Monocyte nonspecific esterase: purification and subunit structure

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 479-487
Author(s):  
J Yourno
Keyword(s):  
Gel Electrophoresis ◽  
Cultured Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Nonspecific Esterase ◽  
Subunit Structure ◽  
Protein Chain ◽  
Cell Extracts

Monocyte nonspecific esterase has been purified from cultured cells of the acute myeloid leukemia cell line, ML-1. The purified enzyme shows the characteristic properties of the monocyte neutral serine carboxyl esterase, with high sensitivity to organophosphorus inhibitors and sodium fluoride inhibitor. The enzyme is a membrane protein which in the native state exists as a monomer of a mol wt of approximately 68,000 and a trimer of mol wt 205,000. These forms exhibit a complex pattern of dissociation and reassociation based on apparent noncovalent binding of subunits. The delipidated dissociated enzyme runs as a single protein chain of a mol wt of approximately 62,000 on sodium dodecyl sulfate (SDS) gel electrophoresis. The relation of the subunits to monocyte isoenzymes seen on isoelectric focusing (IEF) and polyacrylamide gel electrophoresis at pH 9.5 (pH 9.5 PAGE) of cell extracts is demonstrated. Availability of purified enzyme allows development of monoclonal antibodies and analysis of myeloid differentiation. In addition, the substrate specificity and function of the purified monocyte ectoenzyme are being examined.

Characteristics of insulin receptors and insulin action in human myelogenous leukemia cell line K-562

Diabetes ◽  
1985 ◽  
Vol 34 (4) ◽  
pp. 347-352 ◽  
Author(s):  
T. Yamanouchi ◽  
T. Tsushima ◽  
Y. Akanuma ◽  
M. Kasuga ◽  
H. Mizoguchi ◽  
...  
Keyword(s):  
Cell Line ◽  
Insulin Action ◽  
Leukemia Cell ◽  
Insulin Receptors ◽  
Myelogenous Leukemia ◽  
Leukemia Cell Line

Antioxidant effects and in vitro cytotoxicity on human cancer cell lines of flavonoid-rich Flamboyant (Delonix regia (Bojer) Raf.) flower extract

2020 ◽  
Vol 21 ◽  
Author(s):  
Putthiporn Khongkaew ◽  
Phanphen Wattanaarsakit ◽  
Konstantinos I. Papadopoulos ◽  
Watcharaphong Chaemsawang
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Flower Extract ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Murine Leukemia Cell ◽  
Dose Dependent

Background: Cancer is a noncommunicable disease with increasing incidence and mortality rates both worldwide and in Thailand. Its apparent lack of effective treatments is posing challenging public health issues. Introduction: Encouraging research results indicating probable anti-cancer properties of the Delonix regia flower extract (DRE) have prompted us to evaluate the feasibility of developing a type of product for future cancer prevention or treatment. Methods and Results: In the present report, using High Performance Liquid Chromatography (HPLC), we demonstrate in the DRE, the presence of high concentrations of three identifiable flavonoids, namely rutin 4.15±0.30 % w/w, isoquercitrin 3.04±0.02 %w/w, and myricetin 2.61±0.01 % w/w respectively while the IC50 of DPPH and ABTS assay antioxidation activity was 66.88±6.30 µg/ml and 53.65±7.24 µg/ml respectively. Discussion: Our cancer cell line studies using the MTT assay demonstrated DREs potent and dose dependent inhibition of murine leukemia cell line (P-388: 35.28±4.07% of cell viability remaining), as well as of human breast adenocarcinoma (MCF-7), human cervical carcinoma (HeLa), human oral cavity carcinoma (KB), and human colon carcinoma (HT-29) cell lines in that order of magnitude. Conclusion: Three identifiable flavonoids (rutin, isoquercitrin and myricetin) with high antioxidation activity and potent and dose dependent inhibition of murine leukemia cell line and five other cancer cell lines were documented in the DRE. The extract’s lack of cytotoxicity in 3 normal cell lines is a rare advantage not usually seen in current antineoplastic agents. Yet another challenge of the DRE was its low dissolution rate and long-term storage stability, issues to be resolved before a future product can be formulated.

scholarly journals Human Platelet Antigen Genotyping and Expression of CD109 (Human Platelet Antigen 15) mRNA in Various Human Cell Types

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Sang Mee Hwang ◽  
Mi Jung Kim ◽  
Ho Eun Chang ◽  
Yun Ji Hong ◽  
Taek Soo Kim ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Human Platelet ◽  
Human Fibroblasts ◽  
Leukemia Cell ◽  
Embryonic Stem ◽  
Melting Curve ◽  
Leukemia Cell Line ◽  
Melting Curve Analysis ◽  
Platelet Antigen ◽  
Human Platelet Antigen

CD109 gene encodes a glycosylphosphatidylinositol-linked glycoprotein found in a subset of platelets and endothelial cell, and human platelet antigen (HPA) 15 is found on CD109. We evaluated the HPA genotype and/or the CD109 mRNA expression on two peripheral blood stem cells (PBSC), two peripheral bloods (PB), 12 granulocyte products, natural killer (NK)-92, B-lymphocyte (CO88BV59-1), K-562 leukemia cell line, human embryonic stem cell (hESC), and human fibroblasts (HF). HPA genotyping was performed by SNaPshot assay and CD109 mRNA expression was evaluated by real-time PCR with SYBR green and melting curve analysis. Genotype HPA-15a/-15a was found in PBSC#1 and two granulocyte products, and HPA-15a/-15b was found in PBSC#2, eight granulocyte products, NK-92, K-562, hESC, and HF, and HPA-15b/-15b was found in two granulocyte products. CD109 mRNA expression was highly increased in HF and increased in CD34+ and CD34− PBSCs and some granulocyte products, compared to the PB. However, the increase of expression level varied among the PBSC and granulocyte products. The CD109 mRNA expression of NK-92, K-562, hESC, and CO 88BV59-1 was not detected. HPA genotype was evaluated in various cells and the expression of CD109, which contains HPA 15, was different among cell lines and high in HF and PBSCs.

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