A data-driven approach to establishing cell motility patterns as predictors of macrophage subtypes and their relation to cell morphology

The motility of macrophages in response to microenvironment stimuli is a hallmark of innate immunity, where macrophages play pro-inflammatory or pro-reparatory roles depending on their activation status during wound healing. Cell size and shape have been informative in defining macrophage subtypes. Studies show pro- and anti-inflammatory macrophages exhibit distinct migratory behaviors, in vitro, in 3D, and *in vivo, *but this link has not been rigorously studied. We apply both morphology and motility-based image processing approaches to analyze live cell images consisting of macrophage phenotypes. Macrophage subtypes are differentiated from primary murine bone marrow-derived macrophages using a potent lipopolysaccharide (LPS) or cytokine interleukin-4 (IL-4). We show that morphology is tightly linked to motility, which leads to our hypothesis that motility analysis could be used alone or in conjunction with morphological features for improved prediction of macrophage subtypes. We ..., • Macrophage Differentiation: Naïve BMDMs (M0) were differentiated into M1 or M2 subtypes. For M1 differentiation, 100 ng/ml lipopolysaccharide (LPS) was used. For M2 differentiation, 20 ng/ml recombinant mouse interleukin-4 (IL-4) was employed. • Culturing and Activation: The macrophages were seeded in 24-well tissue culture-treated plates and activated as detailed in the accompanying manuscript. • Imaging: Time-lapse contrast images were captured 48 hours post-stimulation using a Carl Zeiss Observer Z1 inverted microscope with a motorized stage and incubation chamber. Imaging was done using the MetaMorph program with a Retiga R6 CCD camera, under 10× or 20× objective lenses. , , # Data from: A data-driven approach to establishing cell motility patterns as predictors of macrophage subtypes and their relation to cell morphology #### Description of the data and file structure **Time-lapse Images of Murine Macrophages** Overview: This dataset comprises time-lapse images of murine macrophages cultured in vitro. The images are provided in TIFF format. Bone marrow was isolated from the femur bones of C57/BL6 mice to generate bone marrow-derived macrophages (BMDMs). We utilized a total of 20 mice, creating five separate batches of BMDMs, with each batch derived from a mixture of bone marrow from 3-6 mice. These BMDMs were then differentiated into naïve (M0), M1, or M2 macrophages using specific stimuli and conditions. ### Files and variables #### Dataset Details: • Data Collection Period: September 2019 to May 2021. • File Naming Convention: Each file is named according to the Macrophage Type, Time Post-Differentiation, Time Interval, Magnification, and Scale. ...,巨噬细胞响应微环境刺激的运动能力是先天免疫的标志性特征之一。在伤口愈合过程中，巨噬细胞会根据自身激活状态分别发挥促炎或促修复的功能。细胞大小与形态可用于区分巨噬细胞亚型。已有研究表明，促炎与抗炎巨噬细胞在体外、三维环境以及体内均表现出截然不同的迁移行为，但二者之间的关联尚未得到严谨探究。本研究采用基于形态学与运动学的图像处理方法，对涵盖不同巨噬细胞表型的活细胞图像进行分析。我们通过强效脂多糖（lipopolysaccharide, LPS）或细胞因子白细胞介素-4（interleukin-4, IL-4）处理原代小鼠骨髓来源巨噬细胞（bone marrow-derived macrophages, BMDMs），以诱导其分化为不同亚型。本研究证实细胞形态与运动能力紧密相关，据此提出假设：可单独采用运动分析，或结合形态学特征，以更精准地预测巨噬细胞亚型。 • 巨噬细胞分化：将未成熟BMDMs（M0型）诱导分化为M1或M2亚型。M1型分化采用100 ng/ml脂多糖（LPS），M2型分化则使用20 ng/ml重组小鼠白细胞介素-4（IL-4）。 • 培养与激活：将巨噬细胞接种于经组织培养处理的24孔板中，并按照随附手稿中的详细步骤完成激活处理。 • 成像：刺激处理48小时后，采用搭载电动载物台与培养舱的卡尔·蔡司（Carl Zeiss）Observer Z1倒置显微镜采集明场延时图像。成像过程通过MetaMorph软件搭配Retiga R6 CCD相机完成，使用10×或20×物镜镜头。 # 数据来源：基于数据驱动的方法确立细胞迁移模式作为巨噬细胞亚型预测指标及其与细胞形态的关联 #### 数据与文件结构说明 **小鼠巨噬细胞延时图像数据集** 概述： 本数据集包含体外培养的小鼠巨噬细胞延时图像，图像格式为TIFF。研究人员从C57/BL6小鼠的股骨中分离骨髓以制备骨髓来源巨噬细胞（BMDMs），共使用20只小鼠，构建了5批独立的BMDMs样本，每批样本均来自3~6只小鼠的骨髓混合液。随后通过特定刺激与培养条件，将这些BMDMs分化为未成熟型（M0）、M1型或M2型巨噬细胞。 ### 文件与变量 #### 数据集详情 • 数据采集周期：2019年9月至2021年5月。 • 文件命名规则：每个文件的命名依据巨噬细胞类型、分化后时间、时间间隔、放大倍数与标尺信息进行编制。 ...