Microfluidic platform for controlling the differentiation of embryoid bodies

Abstract

Embryonic stem (ES) cells are pluripotent cells, which can differentiate into any cell type. This cell type has often been implicated as an eminent source of renewable cells for tissue regeneration and cellular replacement therapies. Studies on manipulation of the various differentiation pathways have been at the forefront of research. There are many ways in which ES cells can be differentiated. One of the most common techniques is to initiate the development of embryoid bodies (EBs) by in vitro aggregation of ES cells. Thereafter, EBs can be induced to undergo differentiation into various cell lineages. In this article, we present a microfluidic platform using biocompatible materials, which is suitable for culturing EBs. The platform is based on a Y-channel device with two inlets for two different culturing media. An EB is located across both streams. Using the laminar characteristics at low Reynolds number and high Peclet numbers, we have induced cell differentiation on half of the EB while maintaining the other half in un-induced stages. The results prove the potential of using microfluidic technology for manipulation of EBs and ES cells in tissue engineering.

Stem cells in microfluidics

“…This device was able to independently cultivate two halves of an EB in two separate media due to the laminar characteristics at low Reynolds number flow in a microchannel. 136 Thus, cell differentiation was induced on half of the EB, while the other half of the EB was maintained in an uninduced state simultaneously. Similarly, by using a patterned laminar flow, it was also demonstrated that patterned delivery of reagents and dissociation enzymes to selected regions of individual hESC colonies was feasible.…”

“…Differentiation of ESCs is often initiated by the generation of embryoid bodies ͑EBs͒, which are three-dimensional cell aggregates formed by culturing ESCs on a nonadherent substrate. 113,114 In this section, a variety of methods for studying the response of ESCs in microfluidic systems are introduced and reviewed, including microwells, [115][116][117][118][119][120][121] membrane fabrication, [122][123][124] feederlayer coating, [125][126][127] arrayed microreactors with microwells, [128][129][130][131] and physical and chemical stimulation 111,[132][133][134][135][136][137][138][139] to study the morphology of ESCs.…”

Engineering Strategies for the Formation of Embryoid Bodies from Human Pluripotent Stem Cells

“…In the study of Fung et al [76], a Y-channel microfluidic device that generates parallel laminar flows of two different culture media without major intermixing has been used to subdifferentiate a single EB into more than one lineage at the same time. By placing EBs across both streams of two separate culture media resulting from laminar coflow in a microchannel, a single EB may be simultaneously induced into different lineages, or control differentiated in some areas, while remaining in undifferentiated stages in the rest of the areas.…”

Induction of alternative fate other than default neuronal fate of embryonic stem cells in a membrane-based two-chambered microbioreactor by cell-secreted BMP4

“…These capabilities of the micro-scale culture systems provide opportunity to study influence of cellsecreted soluble factors on cell behavior in an in vivo relevant dimension and manner. However, previous microfabrication based studies primarily focused on flow 7,8 and size (diameter of ESC aggregates i.e., embryoid bodies (EBs)) 9,10 dependent modulation of soluble microenvironment to direct ESC differentiation. No study focused on the influence of cell-secreted factors in a small culture volume, which might strongly influence ESC differentiation.…”