Effect of spheroid size on gene expression profiles of a mouse mesenchymal stem cell line in spheroid culture
Effect of spheroid size on gene expression profiles of a mouse mesenchymal stem cell line in spheroid culture
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DOI：
10.3233/bme-221406
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发表时间：
2023-01-01
影响因子：
1
通讯作者：
Uchida, Kentaro
中科院分区：
文献类型：
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作者：
Kuroda, Akiyoshi;Mineo, Ayumi;Uchida, Kentaro
背景：基于间充质干细胞（MSC）的疗法提供了骨修复的潜力。通过增加营养因子的分泌，MSC 球体培养物可能比 MSC 单层细胞具有更强的治疗潜力。然而，球体大小对营养因子表达的影响尚不清楚。目的：我们研究球体大小对营养因子相关基因表达的影响。方法：使用小鼠MSC系KUM10。 RNA-seq 用于筛选 MSC 单层和球状培养物的转录谱。通过 q-PCR 在 5 x 10(4)（S 组）、5 x 10(5)（M 组）、5 x 10(6)（L 组）细胞/孔的培养物中评估 RNA-seq 中鉴定的差异表达基因。 结果：比较 KUM10 单层和球状培养物之间的表达水平，鉴定出 2140 个差异表达基因，其中 1047 个上调，1093 个下调。 KUM10 球体。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，L 组中 Fgf9、Fgf18、Vegfa 和 Vegfc 的表达升高，S 组中 Pdgfb 和 Tgfb1 的表达升高。结论：球体大小可能影响营养因子表达。我们的结果将有助于未来研究评估 MSC 球体治疗骨损伤的效用。
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 x 10(4) (S group), 5 x 10(5) (M group), 5 x 10(6) (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.