Single Cell RNAseq REX-1 RFP mESC 2019

This experiment examines how increasing stress (sorbitol) can override normal stemness (NS) conditions, which LIF maintains, causing lineage imbalance and otherwise affecting cells in the early embryo. It also compares NS to normal differentiation (ND) (without LIF). D3 are from 8 blastocysts of 129/Sv+/+ strain mice germ line competent 1. No sex is given as these may actually be a product of different sexes of the 8 blastocysts. The D3 cells were obtained for ATCC (American Type Culture Collection). 1. Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol 1985;87:27-45. Rex1 promoter- Red fluorescence protein (RFP) transgenic viable reporter ESC were created and validated previously2. 2. Li Q, Gomez-Lopez N, Drewlo S, et al. Development and Validation of a Rex1-RFP Potency Activity Reporter Assay That Quantifies Stress-Forced Potency Loss in Mouse Embryonic Stem Cells. Stem Cells Dev 2016;25:320-8. Overall design: Single Cell RNAseq of Rex1 promoter- Red Fluorescent Protein (RFP) mESC cultured for 72hr with different stimuli. Single replicates.