Three-Dimensional Printed Stamps for the Fabrication of Patterned Microwells and High-Throughput Production of Homogeneous Cell Spheroids

Tomas Gonzalez-Fernandez, Alejandro J. Tenorio, J. Kent Leach, J. Kent Leach

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


Aggregation of cells into spheroids and organoids is a promising tool for regenerative medicine, cancer and cell biology, and drug discovery due to their recapitulation of the cell-cell and cell-matrix interactions found in vivo. Traditional approaches for the production of spheroids, such as the hanging drop method, are limited by the lack of reproducibility and the use of labor-intensive and time-consuming techniques. The need for high-throughput approaches allowing for the quick and reproducible formation of cell aggregates has driven the development of soft lithography techniques based on the patterning of microwells into nonadherent hydrogels. However, these methods are also limited by costly, labor-intensive, and multistep protocols that could impact the sterility of the process and efficiency of spheroid formation. In this study, we describe a one-step method for the fabrication of patterned nonadherent microwells into tissue culture plates using three-dimensional (3D) printed stamps and evaluate the production of cell spheroids of different sizes and cell sources. The generation of bone marrow-derived mesenchymal stromal cell and endothelial cell spheroids by the use of 3D printed stamps was superior in comparison with a widely used multistep mold technique, yielding spheroids of larger sizes and higher DNA content. The 3D stamps produced spheroids of more consistent diameter and DNA content when compared with other commercially available methods. These 3D printed stamps offer a tunable, simple, fast, and cost-effective approach for the production of reproducible spheroids and organoids for a wide range of applications.

Original languageEnglish (US)
Pages (from-to)139-147
Number of pages9
Journal3D Printing and Additive Manufacturing
Issue number3
StatePublished - Jun 1 2020


  • 3D cell culture methods
  • 3D printing
  • cell spheroids
  • micropatterning
  • microwell

ASJC Scopus subject areas

  • Materials Science (miscellaneous)
  • Industrial and Manufacturing Engineering


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