User-defined morphogen patterning for directing human cell fate stratification

Sci Rep. 2019 Apr 23;9(1):6433. doi: 10.1038/s41598-019-42874-8.


Concentration gradients of biochemical stimuli such as morphogens play a critical role in directing cell fate patterning across species and throughout development but are not commonly recapitulated in vitro. While in vitro biomolecule gradients have been generated using customized microfluidic platforms, broad implementation has been limited because these platforms introduce new variables to cell culture such as externally driven flow, culture in a specialized matrix, or extended time for in situ long range diffusion. Here we introduce a method that enables preforming and then transferring user-controlled gradients to cells in standard "open" cultures. Our gradient patterning devices are modular and decoupled from the culture substrate. We find that gradient generation and transfer are predictable by finite element modeling and that device and loading parameters can be used to tune the stimulus pattern. Furthermore, we demonstrate use of these devices to spatially define morphogen signal gradients and direct peri-gastrulation fate stratification of human pluripotent stem cells. This method for extrinsic application of biochemical signal gradients can thus be used to spatially influence cellular fate decisions in a user-controlled manner.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Body Patterning / physiology*
  • Cell Culture Techniques*
  • Cell Differentiation
  • Cell Line
  • Collagen / chemistry
  • Drug Combinations
  • Finite Element Analysis
  • Gastrulation / physiology
  • Human Umbilical Vein Endothelial Cells / cytology*
  • Human Umbilical Vein Endothelial Cells / physiology
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / physiology
  • Lab-On-A-Chip Devices
  • Laminin / chemistry
  • Proteoglycans / chemistry
  • Signal Transduction / physiology*


  • Drug Combinations
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen