Automated Fabrication of Streptavidin-Based Self-assembled Materials for High-Content Analysis of Cellular Response to Growth Factors

Abstract

The automation of liquid-handling routines offers great potential for fast, reproducible, and labor-reduced biomaterial fabrication but also requires the development of special protocols. Competitive systems demand for a high degree in miniaturization and parallelization while maintaining flexibility regarding the experimental design. Today, there are only a few possibilities for automated fabrication of biomaterials inside multiwell plates. We have previously demonstrated that streptavidin-based biomimetic platforms can be employed to study cellular behaviors on biomimetic surfaces. So far, these self-assembled materials were made by stepwise assembly of the components using manual pipetting. In this work, we introduce for the first time a fully automated and adaptable workflow to functionalize glass-bottom multiwell plates with customized biomimetic platforms deposited in single wells using a liquid-handling robot. We then characterize the cell response using automated image acquisition and subsequent analysis. Furthermore, the molecular surface density of the biomimetic platforms was characterized in situ using fluorescence-based image correlation spectroscopy. These measurements were in agreement with standard ex situ spectroscopic ellipsometry measurements. Due to automation, we could do a proof of concept to study the effect of heparan sulfate on the bioactivity of bone morphogenetic proteins on myoblast cells, using four different bone morphogenetic proteins (BMPs) (2, 4, 6, and 7) in parallel, at five increasing concentrations. Using such an automated self-assembly of biomimetic materials, it may be envisioned to further investigate the role of a large variety of extracellular matrix (ECM) components and growth factors on cell signaling.

Sweet but Challenging: Tackling the Complexity of GAGs with Engineered Tailor‐Made Biomaterials

“…Adapted with permission. [34] Copyright 2022, American Chemical Society. chemokines CCL2, CCL5, CCL7, CCL13, CXCL8, and CXCL10 to 47 synthesized tetrasaccharides.…”

“…More recently, the automation of the fabrication of self‐assembled materials in the form of streptavidin‐based materials has been developed. [ 34 ] Such materials deposited in 96‐well cell culture microplates can be used for high‐content studies of cellular responses. Notably, cells can be cultured on GAG‐based biomaterials deposited directly at the bottom of the microplate, and can be stimulated by growth factors adsorbed onto the GAGs (Figure 7B).…”

“…Automated strategies for biomaterial fabrication and cell behavior analysis have recently emerged. [33,34] We anticipate that novel high-content methodologies for the study of GAG-GF interactions and the role of GAGs mediated presentation of GFs on cellular responses will emerge and will be further developed.…”

“…A dose-response curve of nuclear pSMAD1/5/9 was established for all BMPs bound to HS. Adapted with permission [34]. Copyright 2022, American Chemical Society.…”

Automated, High-Throughput Screening of Hybrid Elastin-like Polypeptide/Polysaccharide Multilayer Film Deposition

“…Notably, the LbL method traditionally relies on automated approaches for the self-assembly of the multilayers on the target surface via dipping, spraying, and spinning methods, which limit the number of film formulations tested. The preliminary LbL high-throughput approaches previously described , and used to screen soft materials , also present limitations regarding both the technology affordability and the protocol access and versatility. As a result, studies involving LbL-based films restrict their investigation to a handful of experimental conditions to efficiently describe the relevant features in materials’ performance. ,, The assessment of a broader experimental space in these circumstances might lead to an in-depth understanding and optimal performance of the investigated system.…”