| Speaker: |
Micha Drukker
Stanford Institute for Stem Cell Biology |
| Title: |
Prospective Isolation of Blastocyst- and Epiblast-Stage Precursors from Human Embryonic Stem Cells |
| Abstract : | To explore the landscape and properties of the earliest precursors derived from hESCs we conducted a flow cytometry screen with over 400 monoclonal antibodies against cell surface markers. This screen identified distinct subsets of putative precursors that are either present in undifferentiated cultures, or that emerge following 3 days of differentiation in response to BMP4 and retinoic acid. By isolating over 30 cell subpopulations and analyzing the expression of developmental genes, we identified 4 distinct candidate precursor populations corresponding respectively, to embryonic primitive endoderm of the pre-gastrulation stage and to gastrulation-stage precursors of the visceral streak (meso-endoderm), allantois and chorion. Single cell analysis revealed a surprising transcriptional state of the visceral endoderm-like cells, characterized by strong induction of a wide range of differentiation associated genes but only a relatively marginal decrease in pluripotency genes. In addition, a small portion of the meso-endoderm-like cells co-expressed pan-mesoderm and endoderm genes, suggesting the existence of human bi-potent meso-endoderm precursors. Prospective isolation of cell populations with striking similarities to pre-gastrulation and gastrulation-stage precursors provides an important platform for studying human development with unprecedented resolution, and may facilitate production of tissue progenitors for therapeutic applications from hES and induced pluripotent cell lines. |
