Effect of spheroid size on gene expression profiles of a mouse mesenchymal stem cell line in spheroid culture.

IF 1 4区医学 Q4 ENGINEERING, BIOMEDICAL

Bio-medical materials and engineering Pub Date : 2023-01-01 DOI:10.3233/BME-221406

Akiyoshi Kuroda, Ayumi Mineo, Shintaro Shoji, Gen Inoue, Wataru Saito, Hiroyuki Sekiguchi, Masashi Takaso, Kentaro Uchida

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引用次数: 2

Abstract

Background: Mesenchymal stem cell (MSC)-based therapies offer potential for bone repair. MSC spheroid cultures may harbor enhanced therapeutic potential over MSC monolayers through increased secretion of trophic factors. However, the impact of spheroid size on trophic factor expression is unclear.

Objective: We investigated the effect of spheroid size on trophic factor-related gene expression.

Methods: KUM10, a murine MSC line was used. RNA-seq was used to screen the transcriptional profiles of MSC monolayer and spheroid cultures. Differentially expressed genes identified in RNA-seq were evaluated by q-PCR in cultures of 5 × 104 (S group), 5 × 105 (M group), 5 × 106 (L group) cells/well.

Results: Comparison of expression levels between KUM10 monolayer and spheroid cultures identified 2140 differentially expressed genes, of which 1047 were upregulated and 1093 were downregulated in KUM10 spheroids. Among these, 12 upregulated genes (Bmp2, Fgf9, Fgf18, Ngf, Pdgfa, Pdgfb, Tgfb1, Vegfa, Vegfc, Wnt4, Wnt5a, Wnt10a) were associated with secretory growth factors. Of these, expression of Fgf9, Fgf18, Vegfa and Vegfc was elevated in the L group, and Pdgfb and Tgfb1 was elevated in the S group.

Conclusions: Spheroid size may impact trophic factor expression. Our results will be useful for future studies assessing the utility of MSC spheroids for treating bone injury.

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球形细胞大小对小鼠间充质干细胞基因表达谱的影响。

背景:间充质干细胞(MSC)为骨修复提供了潜在的治疗方法。通过增加营养因子的分泌，骨髓间充质干细胞球形培养物可能比骨髓间充质干细胞单层培养物具有更高的治疗潜力。然而，球体大小对营养因子表达的影响尚不清楚。目的:研究球体大小对营养因子相关基因表达的影响。方法:采用小鼠MSC细胞系KUM10。RNA-seq用于筛选MSC单层和球形培养的转录谱。在5 × 104 (S组)、5 × 105 (M组)、5 × 106 (L组)细胞/孔的培养中，采用q-PCR检测RNA-seq鉴定的差异表达基因。结果:通过KUM10单层培养与球状培养的表达水平比较，鉴定出2140个差异表达基因，其中1047个表达上调，1093个表达下调。其中，12个上调基因(Bmp2、Fgf9、Fgf18、Ngf、Pdgfa、Pdgfb、Tgfb1、Vegfa、Vegfc、Wnt4、Wnt5a、Wnt10a)与分泌性生长因子相关。其中，Fgf9、Fgf18、Vegfa、Vegfc在L组表达升高，Pdgfb、Tgfb1在S组表达升高。结论:球体大小可能影响营养因子的表达。我们的结果将对未来评估MSC球体治疗骨损伤的效用的研究有用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bio-medical materials and engineering 工程技术-材料科学：生物材料

CiteScore

1.80

自引率

0.00%

发文量

审稿时长

6 months

期刊介绍： The aim of Bio-Medical Materials and Engineering is to promote the welfare of humans and to help them keep healthy. This international journal is an interdisciplinary journal that publishes original research papers, review articles and brief notes on materials and engineering for biological and medical systems. Articles in this peer-reviewed journal cover a wide range of topics, including, but not limited to: Engineering as applied to improving diagnosis, therapy, and prevention of disease and injury, and better substitutes for damaged or disabled human organs; Studies of biomaterial interactions with the human body, bio-compatibility, interfacial and interaction problems; Biomechanical behavior under biological and/or medical conditions; Mechanical and biological properties of membrane biomaterials; Cellular and tissue engineering, physiological, biophysical, biochemical bioengineering aspects; Implant failure fields and degradation of implants. Biomimetics engineering and materials including system analysis as supporter for aged people and as rehabilitation; Bioengineering and materials technology as applied to the decontamination against environmental problems; Biosensors, bioreactors, bioprocess instrumentation and control system; Application to food engineering; Standardization problems on biomaterials and related products; Assessment of reliability and safety of biomedical materials and man-machine systems; and Product liability of biomaterials and related products.