Determination of adipogenesis stages of human umbilical cord-derived mesenchymal stem cells using three-dimensional label-free holotomography

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods Pub Date : 2024-10-11 DOI:10.1016/j.ymeth.2024.10.005

Mahesh Prakash Bhatta , Gun-Woo Won , Seung Hoon Lee , Seung-Hyeon Choi , Cheong-Hae Oh , Ji Hyun Moon , Hong-Hoa Hoang , Jaehyeok Lee , Sang Do Lee , Jong-Il Park

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引用次数: 0

Abstract

Adipogenesis involves complex changes in gene expression, morphology, and cytoskeletal organization. However, the quantitative analysis of live cell images to identify their stages through morphological markers is limited. Distinct adipogenesis markers on human umbilical cord-derived mesenchymal stem cells (UC-MSCs) were identified through holotomography, a label-free live cell imaging technique. In the MSC-to-preadipocyte transition, the nucleus-to-cytoplasm ratio (0.080 vs. 0.052) and lipid droplet (LD) refractive index variation decreased (0.149 % vs. 0.061 %), whereas the LD number (20 vs. 65) increased. This event was also accompanied by the downregulation and upregulation of THY1 and Preadipocyte Factor-1 (PREF-1), respectively. In the preadipocyte to immature adipocyte shift, cell sphericity (0.20 vs. 0.43) and LD number (65 vs. 200) surged, large LDs (>10 μm³) appeared, and the major axis of the cell was reduced (143.7 μm vs. 83.12 μm). These findings indicate features of preadipocyte and immature adipocyte stages, alongside the downregulation of PREF-1 and upregulation of Peroxisome Proliferator-Activated Receptor gamma (PPARγ). In adipocyte maturation, along with PPARγ and Fatty Acid-Binding Protein 4 upregulation, cell compactness (0.15 vs. 0.29) and sphericity (0.43 vs. 0.59) increased, and larger LDs (>30 μm³) formed, marking immature and mature adipocyte stages. The study highlights the distinct adipogenic morphological biomarkers of adipogenesis stages in UC-MSCs, providing potential applications in biomedical and clinical settings, such as fostering innovative medical strategies for treating metabolic disease.

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利用三维无标记全息图确定人脐带间充质干细胞的脂肪生成阶段

脂肪生成涉及基因表达、形态和细胞骨架组织的复杂变化。然而，通过形态标记对活细胞图像进行定量分析以确定其阶段的方法还很有限。通过无标记活细胞成像技术--全息图像技术，确定了人脐带间充质干细胞（UC-MSCs）上不同的脂肪生成标记。在间充质干细胞向脂肪细胞转变的过程中，细胞核与细胞质的比率（0.080 对 0.052）和脂滴（LD）折射率变化降低（0.149 % 对 0.061 %），而脂滴数量增加（20 对 65）。与此同时，THY1 和前脂肪细胞因子-1（PREF-1）也分别出现了下调和上调。在前脂肪细胞向未成熟脂肪细胞转变的过程中，细胞球形度（0.20 对 0.43）和 LD 数量（65 对 200）激增，出现大的 LD（10 μm3），细胞主轴缩小（143.7 μm 对 83.12 μm）。这些发现显示了前脂肪细胞和未成熟脂肪细胞阶段的特征，以及 PREF-1 的下调和过氧化物酶体激活受体γ（PPARγ）的上调。在脂肪细胞成熟过程中，随着 PPARγ 和脂肪酸结合蛋白 4 的上调，细胞紧密度（0.15 vs. 0.29）和球形度（0.43 vs. 0.59）增加，形成更大的 LD（30 μm3），标志着脂肪细胞的未成熟和成熟阶段。该研究强调了 UC 间充质干细胞中不同脂肪生成阶段的脂肪生成形态生物标志物，为生物医学和临床提供了潜在的应用前景，如促进治疗代谢性疾病的创新医疗策略。

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来源期刊

Methods 生物-生化研究方法

CiteScore

9.80

自引率

2.10%

发文量

222

审稿时长

11.3 weeks

期刊介绍： Methods focuses on rapidly developing techniques in the experimental biological and medical sciences. Each topical issue, organized by a guest editor who is an expert in the area covered, consists solely of invited quality articles by specialist authors, many of them reviews. Issues are devoted to specific technical approaches with emphasis on clear detailed descriptions of protocols that allow them to be reproduced easily. The background information provided enables researchers to understand the principles underlying the methods; other helpful sections include comparisons of alternative methods giving the advantages and disadvantages of particular methods, guidance on avoiding potential pitfalls, and suggestions for troubleshooting.