Generation of 3D Soluble Signal Gradients in Cell‐Laden Hydrogels Using Passive Diffusion

• DOI: 10.1002/adbi.201800237
• Publication Date: 2018-11-15T11:51:07Z
• Authors: Lucas Ahrens; Simon Tanaka; Daniel Vonwil; Jon Christensen; Dagmar Iber; V. Prasad Shastri
• Keywords: SIGNAL (programming language)

Soluble signal gradients play an important role in organ patterning, cell migration, and differentiation. Currently, 2D culture are realized using microfluidics here cells exposed to high nonphysiological shear stress. Tissue morphogenesis (organogenesis) however occurs 3D therefore there is a need for simple practical systems impose dispersed matrix. Herein, gradient generator based on passive diffusion elements that recapitulates interstitial flow capable of imposing predictable over long length scales (6 mm) lasting up 48 h hydrogel environment reported. Using recombinant human WNT3A (rhWNT3A), the spatiotemporal activation canonical WNT pathway epithelial kidney mesenchymal stems expressing green fluorescence protein reporter transcription factor/lymphoid enhancer-binding factor (TCF/LEF) promoter demonstrated. By refining computation models experimental findings, coefficient rhWNT3A presence determined. Furthermore, formation rhBMP4 visualized immunohistochemistry by staining phospho-SMAD1/5, downstream targets bone morphogenetic (BMP) pathway. The simplicity expected spur its adoption studying developmental biology paradigms vitro.