title: High-throughput fingerprinting of human pluripotent stem cell fate responses and lineage bias.
abstract: Populations of cells create local environments that lead to emergent heterogeneity. This is particularly evident with human pluripotent stem cells (hPSCs): microenvironmental heterogeneity limits hPSC cell fate control. We developed a high-throughput platform to screen hPSCs in configurable microenvironments in which we optimized colony size, cell density and other parameters to achieve rapid and robust cell fate responses to exogenous cues. We used this platform to perform single-cell protein expression profiling, revealing that Oct4 and Sox2 costaining discriminates pluripotent, neuroectoderm, primitive streak and extraembryonic cell fates. We applied this Oct4-Sox2 code to analyze dose responses of 27 developmental factors to obtain lineage-specific concentration optima and to quantify cell line-specific endogenous signaling pathway activation and differentiation bias. We demonstrated that short-term responses predict definitive endoderm induction efficiency and can be used to rescue differentiation of cell lines reticent to cardiac induction. This platform will facilitate high-throughput hPSC-based screening and quantification of lineage-induction bias.
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Emanuel J P
authors.initials: EJ
authors.lastname: Nazareth
authors.forename: Joel E E
authors.initials: JE
authors.lastname: Ostblom
authors.forename: Petra B
authors.initials: PB
authors.lastname: Lücker
authors.forename: Shreya
authors.initials: S
authors.lastname: Shukla
authors.forename: Manuel M
authors.initials: MM
authors.lastname: Alvarez
authors.forename: Steve K W
authors.initials: SK
authors.lastname: Oh
authors.forename: Ting
authors.initials: T
authors.lastname: Yin
authors.forename: Peter W
authors.initials: PW
authors.lastname: Zandstra
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D017077:Culture Media, Conditioned / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology; D004707:Endoderm / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology*; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors
chemical_list: D017077:Culture Media, Conditioned; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors