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Purpose <p>3D bioprinting of adipose mesenchymal stromal cells (ASC), as single cells or spheroids, underpins the development of experimental biodressing and secretome-oriented workflows. The assessment of viability-associated readouts is technically challenging in hydrogels. We aimed to evaluate the applicability and comparative performance of CellTiter Glo 3D (CTG-3D) and resazurin in a defined ASC-laden gelatin-alginate architecture intended for downstream workflows in 3D bioprinted constructs.</p> Methods <p>MSCs from C57BL/6 adipose tissue (P3) were characterized and incorporated as free cells into a gelatin–alginate bioink, bioprinted and cultured in DMEM-F12 with or without 10% FBS. After 48&#xa0;h, punched fragments were analyzed with CTG-3D and Resazurin following manufacturer instructions. Luminescence and absorbance were read on an EnVision system.</p> Results <p>Both assays detected the expected increase in viability-associated readouts in FBS-rich medium. CTG-3D showed statistical sensitivity (<i>p</i> = 0.0039) comparable with Resazurin (<i>p</i> = 0.0013). The similarity between assays likely reflects the low-density hydrogel, permitting adequate reagent diffusion and viability-related signal generation.</p> Conclusion <p>The gelatin–alginate bioprinted model supports short-term MSC viability-associated readouts and is suitable for refining culture parameters and bioactive factor release toward experimental biodressing-oriented workflows. CTG-3D and Resazurin perform comparably in this architecture, but assay choice should balance cost and sensitivity according to construct density and complexity.</p> Graphical Abstract <p></p>

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Comparative performance of CTG-3D and resazurin assays in low-density gelatin-alginate bioprinted mesenchymal stromal cell constructs

  • 期刊论文

  • 发表日期：

    2026年6月16日

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  • Isabela Souza Marques,
  • Maria Eduarda Oliveira Borges,
  • Manuela Abrantes do Rego,
  • Karina Ribeiro Silva Pereira,
  • Genilza Pereira de Oliveira,
  • Débora dos Santos Ornellas,
  • Erika Afonso Costa Cortez Marques,
  • Alessandra Alves Thole,
  • Simone Nunes de Carvalho

摘要

Purpose

3D bioprinting of adipose mesenchymal stromal cells (ASC), as single cells or spheroids, underpins the development of experimental biodressing and secretome-oriented workflows. The assessment of viability-associated readouts is technically challenging in hydrogels. We aimed to evaluate the applicability and comparative performance of CellTiter Glo 3D (CTG-3D) and resazurin in a defined ASC-laden gelatin-alginate architecture intended for downstream workflows in 3D bioprinted constructs.

Methods

MSCs from C57BL/6 adipose tissue (P3) were characterized and incorporated as free cells into a gelatin–alginate bioink, bioprinted and cultured in DMEM-F12 with or without 10% FBS. After 48 h, punched fragments were analyzed with CTG-3D and Resazurin following manufacturer instructions. Luminescence and absorbance were read on an EnVision system.

Results

Both assays detected the expected increase in viability-associated readouts in FBS-rich medium. CTG-3D showed statistical sensitivity (p = 0.0039) comparable with Resazurin (p = 0.0013). The similarity between assays likely reflects the low-density hydrogel, permitting adequate reagent diffusion and viability-related signal generation.

Conclusion

The gelatin–alginate bioprinted model supports short-term MSC viability-associated readouts and is suitable for refining culture parameters and bioactive factor release toward experimental biodressing-oriented workflows. CTG-3D and Resazurin perform comparably in this architecture, but assay choice should balance cost and sensitivity according to construct density and complexity.

Graphical Abstract