We have developed a miniaturized 3D stem cell culture array on a functionalized glass slide for spatially addressable high-throughput screening. A microarray spotter was used to deposit cells onto a modified glass surface to yield an array consisting of cells encapsulated in alginate gel spots with volumes as low as 60 nL. A method based on an immunofluorescence technique scaled down to function on a cellular microarray was also used to quantify specific cell marker protein levels in situ. Our results revealed that this platform is suitable for studying the expansion and neural commitment of mouse embryonic stem (ES) cells as they retain their pluripotent and undifferentiated state, and are able to generate neuroectodermal precursor cells characterized by expression of the neural marker Sox-1. In addition, the high-throughput capacity of the platform was tested using a dual-slide system that allowed rapid screening of the effects of tretinoin and fibroblast growth factor-4 (FGF-4) on the pluripotency of mouse ES cells. This high-throughput platform may represent a powerful new tool for investigating cellular mechanisms involved in stem cell expansion and differentiation, and provides the basis for rapid identification of signals and conditions that can be used to direct cellular responses.

3D cell culture microarray for high-throughput studies of stem cell fate

241st National Meeting (2011)

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