Fibroblast Derived Human Engineered Connective Tissue for Screening Applications

Gabriela L. Santos; Tim Meyer; Malte Tiburcy; Alisa DeGrave; Wolfram-Hubertus Zimmermann; Susanne Lutz

doi:10.3791/62700

Fibroblast Derived Human Engineered Connective Tissue for Screening Applications

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Fibroblast Derived Human Engineered Connective Tissue for Screening Applications

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09:50 min

August 20, 2021

DOI: 10.3791/62700-v

Gabriela L. Santos^1,2, Tim Meyer^1,2, Malte Tiburcy^1,2, Alisa DeGrave^1,2, Wolfram-Hubertus Zimmermann^1,2,3,4,5, Susanne Lutz^1,2

¹Institute of Pharmacology and Toxicology,University Medical Center Goettingen, ²DZHK (German Center for Cardiovascular Research) partner site, Goettingen, ³Cluster of Excellence “Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells” (MBExC),University of Goettingen, ⁴Center for Neurodegenerative Diseases (DZNE), ⁵Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP)

Presented here is a protocol to generate engineered connective tissues for a parallel culture of 48 tissues in a multi-well plate with double poles, suitable for mechanistic studies, disease modeling, and screening applications. The protocol is compatible with fibroblasts from different organs and species and is exemplified here with human primary cardiac fibroblasts.

This protocol allows the fibroblast build and organize their own environment under defined mechanical conditions. These results in anisotropic tissues with stiffness and matrix compositions comparable to the cell's natural environment. This method allows to compare up to 48 conditions in parallel.

It is flexible with respect to the used cell type and culture conditions. By using only a few well-defined components, it is reproducible between laboratories. By applying pro-fibrotic factors or antifibrotic drugs, this protocol can be used to study the underlying processes and therapies of fibrotic diseases of any kind.

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