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  • An automated do-it-yourself system for dynamic stem cell and organoid culture in standard multi-well plates.

An automated do-it-yourself system for dynamic stem cell and organoid culture in standard multi-well plates.

Cell reports methods (2022-07-27)
Julia Tischler, Zoe Swank, Hao-An Hsiung, Stefano Vianello, Matthias P Lutolf, Sebastian J Maerkl
ABSTRACT

We present a low-cost, do-it-yourself system for complex mammalian cell culture under dynamically changing medium formulations by integrating conventional multi-well tissue culture plates with simple microfluidic control and system automation. We demonstrate the generation of complex concentration profiles, enabling the investigation of sophisticated input-response relations. We further apply our automated cell-culturing platform to the dynamic stimulation of two widely employed stem-cell-based in vitro models for early mammalian development: the conversion of naive mouse embryonic stem cells into epiblast-like cells and mouse 3D gastruloids. Performing automated medium-switch experiments, we systematically investigate cell fate commitment along the developmental trajectory toward mouse epiblast fate and examine symmetry-breaking, germ layer formation, and cardiac differentiation in mouse 3D gastruloids as a function of time-varying Wnt pathway activation. With these proof-of-principle examples, we demonstrate a highly versatile and scalable tool that can be adapted to specific research questions, experimental demands, and model systems.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Human Plasma Fibronectin Purified Protein, from human plasma, liquid, 1 mg/mL (100 MG pack size is lyophilized), purified protein, suitable for cell culture
Sigma-Aldrich
GSK-3 Inhibitor XVI, GSK-3 Inhibitor XVI - CAS 252917-06-9, is a cell-permeable, potent, ATP-competitive, and highly selective GSK-3 inhibitor (IC₅₀ = 10 and 6.7 nM against GSK-3α and GSK-3β, respectively).