Scholarlink

Background <p>Acoustic fields enable contactless manipulation of cells and multicellular assemblies, offering unique opportunities for three-dimensional (3D) cell culture and mechanobiology. However, standardized and reproducible methodologies that combine sound-guided cell assembly with controlled acoustic stimulation and downstream molecular analysis are still lacking.</p> Methods <p>We developed a reproducible workflow integrating sound-guided assembly and acoustic stimulation of human bone marrow–derived mesenchymal stromal cell (hBMSC) spheroids within 3D fibrin hydrogels. Using a sound-induced morphogenesis (SIM) platform, spheroids were rapidly organized into circular patterns through low-frequency acoustic. Pattern geometry and stability were quantitatively assessed by image-based radial profile analysis. To evaluate biological responsiveness to assembly and stimulation, spheroids were analyzed under three conditions (randomly dispersed, sound-assembled, and sound-assembled with daily acoustic stimulation) followed by transcriptomic profiling using RNA sequencing and validation by RT-qPCR.</p> Results <p>Sound-guided assembly consistently generated stable, high-density annular spheroid patterns that were maintained over culture duration and unaffected by subsequent acoustic stimulation. Transcriptomic analysis revealed reproducible changes in gene expression associated with cell–cell interactions, stress regulation, and mechanosensing in assembled spheroids compared with dispersed controls. Additional acoustic stimulation induced a distinct transcriptional signature involving genes related to matrix remodeling and mechanotransduction, confirming biological sensitivity to applied mechanical cues.</p> Conclusions <p>This method provides a standardized, non-invasive approach to couple spatial cell organization with controlled mechanical stimulation in 3D culture systems. The workflow is compatible with conventional cell culture practices, adaptable to other cell types and hydrogels, and suitable for mechanistic studies investigating how physical cues regulate cellular gene expression. It offers a transferable methodological framework for mechanobiology research and engineered tissue model development.</p> Graphical Abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Control of human mesenchymal stromal cell gene expression levels by sound-guided assembly and stimulation

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月1日

查看全文
  • Elena Della Bella,
  • Farah Daou,
  • Alessandro Cianciosi,
  • Junxuan Ma,
  • Riccardo Tognato,
  • Mauro Alini,
  • Andrea Cochis,
  • Lia Rimondini,
  • Martin J Stoddart,
  • Tiziano Serra

摘要

Background

Acoustic fields enable contactless manipulation of cells and multicellular assemblies, offering unique opportunities for three-dimensional (3D) cell culture and mechanobiology. However, standardized and reproducible methodologies that combine sound-guided cell assembly with controlled acoustic stimulation and downstream molecular analysis are still lacking.

Methods

We developed a reproducible workflow integrating sound-guided assembly and acoustic stimulation of human bone marrow–derived mesenchymal stromal cell (hBMSC) spheroids within 3D fibrin hydrogels. Using a sound-induced morphogenesis (SIM) platform, spheroids were rapidly organized into circular patterns through low-frequency acoustic. Pattern geometry and stability were quantitatively assessed by image-based radial profile analysis. To evaluate biological responsiveness to assembly and stimulation, spheroids were analyzed under three conditions (randomly dispersed, sound-assembled, and sound-assembled with daily acoustic stimulation) followed by transcriptomic profiling using RNA sequencing and validation by RT-qPCR.

Results

Sound-guided assembly consistently generated stable, high-density annular spheroid patterns that were maintained over culture duration and unaffected by subsequent acoustic stimulation. Transcriptomic analysis revealed reproducible changes in gene expression associated with cell–cell interactions, stress regulation, and mechanosensing in assembled spheroids compared with dispersed controls. Additional acoustic stimulation induced a distinct transcriptional signature involving genes related to matrix remodeling and mechanotransduction, confirming biological sensitivity to applied mechanical cues.

Conclusions

This method provides a standardized, non-invasive approach to couple spatial cell organization with controlled mechanical stimulation in 3D culture systems. The workflow is compatible with conventional cell culture practices, adaptable to other cell types and hydrogels, and suitable for mechanistic studies investigating how physical cues regulate cellular gene expression. It offers a transferable methodological framework for mechanobiology research and engineered tissue model development.

Graphical Abstract