title: Software-assisted morphometric and phenotype analyses of human peripheral blood monocyte-derived macrophages induced by a microenvironment model of human esophageal squamous cell carcinoma.
abstract: Human macrophages play important roles in tumor promotion and are called tumor-associated macrophages (TAMs). We previously demonstrated that human esophageal squamous cell carcinomas (ESCCs) contain TAMs and that these TAMs tend to have tumor-supporting features. Here we exposed human macrophages to conditioned media of TE-series human ESCC cell lines (TECMs) to generate an ESCC extracellular stimuli-influenced TAM model. CD14(+) peripheral blood monocytes (PBMos) from healthy donors were treated with M-CSF and with additional IL-4 or TECM exposure. Morphological changes of the cells and the induction of CD163/CD204 proteins were detected in the TECM-exposed model TAMs by immunofluorescence. A software-assisted immunofluorescent cell image analysis showed increased CD163/CD204 positivity in the model TAMs and a weak to moderate positive correlation between the cytoplasmic area and the sum fluorescent intensity of CD204. Morphological changes of the cells were significantly reflected by several cytomorphometric parameters. PBMos were elongated with M-CSF treatment, then enlarged with TECM exposure. The cytoplasmic aspect ratio was decreased by M-CSF treatment and slightly increased by TECM exposure. The nuclear-cytoplasmic ratio decreased during the whole process of cell differentiation. This system is useful for quantitative assessments of TAM-like morphological changes of macrophages and the induction of CD163/CD204 in a model ESCC microenvironment.
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Nishio
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.|Division of Gastro-intestinal Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Urakawa
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.|Oral and Maxillofacial Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Shigeoka
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.|Division of Gastro-intestinal Surgery, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Takase
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Ichihara
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Arai
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Yu-ichiro
authors.initials: Y
authors.lastname: Koma
authors.affiliation: Division of Pathology, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yokozaki
mesh_terms: D015703:Antigens, CD / Q000378:metabolism*; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D017077:Culture Media, Conditioned; D004938:Esophageal Neoplasms / Q000378:metabolism / Q000473:pathology*; D000077277:Esophageal Squamous Cell Carcinoma; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015847:Interleukin-4 / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism / Q000473:pathology*; D009000:Monocytes / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D011956:Receptors, Cell Surface / Q000378:metabolism*; D051117:Scavenger Receptors, Class A / Q000378:metabolism*; D012984:Software; D059016:Tumor Microenvironment; D000098902:CD163 Antigen
chemical_list: D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D000098902:CD163 Antigen; D017077:Culture Media, Conditioned; C508598:IL4 protein, human; C495139:MSR1 protein, human; D011956:Receptors, Cell Surface; D051117:Scavenger Receptors, Class A; D015847:Interleukin-4
keywords: esophageal cancer; image analysis; macrophages; tumor microenvironment