Abstract
Standard tissue culture of adherent cells is known to poorly replicate physiology and often entails suspending cells in solution for analysis and sorting, which modulates protein expression and eliminates intercellular connections. To allow adherent culture and processing in flow, we present 3D-shaped hydrogel cell microcarriers, which are designed with a recessed nook in a first dimension to provide a tunable shear-stress shelter for cell growth, and a dumbbell shape in an orthogonal direction to allow for self-alignment in a confined flow, important for processing in flow and imaging flow cytometry. We designed a method to rapidly design, using the genetic algorithm, and manufacture the microcarriers at scale using a transient liquid molding optofluidic approach. The ability to precisely engineer the microcarriers solves fundamental challenges with shear-stress-induced cell damage during liquid-handling, and is poised to enable adherent cell culture, in-flow analysis, and sorting in a single format.
Highlights
Traditional processes of tissue culture of adherent cells make use of cell growth on flat and rigid polymer petri dishes, flasks, and well plates
Subsequent cell analysis involves scanning the culture surface with microscopy, or bringing cells into suspension with enzymatic or physical treatments followed by flow cytometry to analyze and select sub-populations
We make use of a novel microparticle manufacturing approach called optical transient liquid molding (OTLM), which is capable of producing new classes of complex microparticles with software-designed 3D shape and functionality[13]
Summary
Traditional processes of tissue culture of adherent cells make use of cell growth on flat and rigid polymer petri dishes, flasks, and well plates. Subsequent cell analysis involves scanning the culture surface with microscopy, or bringing cells into suspension with enzymatic or physical treatments followed by flow cytometry to analyze and select sub-populations. This paradigm of cell culture, single-cell enzymatic suspension, and passaging is especially challenging for growth of terminally differentiated cell populations from pluripotent or multipotent precursors[1]. Particle-based cell culture, whereby adherent cells grow and are analyzed on engineered microparticles or microcarriers, can serve as a new paradigm to accelerate culture, passaging, and analysis, without exposing cells to harsh environments[2,3].
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