Heterogeneity within human embryonic stem (hES) cell populations complicates their analysis and clinical use. We have developed a microfluidic cell culture platform consisting of 160 individually addressable 500x500x100 �m chambers adapted to the perfusion culture of isolated hES cell colonies. This device was engineered for the quantification of growth, morphology and transcript expression during maintenance or differentiation. The growth rates and colony morphologies in the device matched cells grown conventionally. Upon induction of differentiation for 6 days using fetal bovine serum, the distribution of clonal populations cultured in the device displayed significant shifts in growth rate (44% decreased), colony morphology (125% increased mean cell area) and OCT4 expression (77% decreased). The change in mean OCT4 expression was revealed to be the result of an emerging bi-modal population distribution. This system provides live-cell imaging and analysis of the dynamics of directed stem cell differentiation, including the ability to resolve clonal differences.

Clonal culture of human embryonic stem cells in a microfluidic device for the analysis of population heterogeneity

245th National Meeting (2013)

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