Micropatterning the organization of multicellular structures in 3D biological hydrogels; insights into collective cellular mechanical interactions

Bar Ergaz, Shahar Goren, Ayelet Lesman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Control over the organization of cells at the microscale level within supporting biomaterials can push forward the construction of complex tissue architectures for tissue engineering applications and enable fundamental studies of how tissue structure relates to its function. While cells patterning on 2D substrates is a relatively established and available procedure, micropatterning cells in biomimetic 3D hydrogels has been more challenging, especially with micro-scale resolution, and currently relies on sophisticated tools and protocols. We present a robust and accessible ‘peel-off’ method to micropattern large arrays of individual cells or cell-clusters of precise sizes in biological 3D hydrogels, such as fibrin and collagen gels, with control over cell-cell separation distance and neighboring cells position. We further demonstrate partial control over cell position in the z-dimension by stacking two layers in varying distances between the layers. To demonstrate the potential of the micropatterning gel platform, we study the matrix-mediated mechanical interaction between array of cells that are accurately separated in defined distances. A collective process of intense cell-generated densified bands emerging in the gel between near neighbors was identified, along which cells preferentially migrate, a process relevant to tissue morphogenesis. The presented 3D gel micropatterning method can be used to reveal fundamental morphogenetic processes, and to reconstruct any tissue geometry with micrometer resolution in 3D biomimetic gel environments, leveraging the engineering of tissues in complex architectures.

Original languageEnglish
Article number015012
Issue number1
StatePublished - 1 Jan 2024


  • cell-matrix interaction
  • fibrous extracellular matrix
  • fibrous hydrogel
  • long-range cell-cell mechanical interaction
  • micropatterning
  • morphogenesis
  • traction force


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