in vitro construction of an embryonic caudal organizer

Purpose: Characterization of cell types in bmp4 explants and BMP4 induced human embryoids. Methods: Single cell transcriptomes of bmp4 explants were harvested at 6 hpf, 8 hpf,10 hpf, 12 hpf, 18 hpf and 24 hpf. Single cell transcriptomes of BMP4 induced human embryoids were harvested at Day 5 and Day 6. Libraries were prepared using Single Cell 3′Library & gel Bead kit v3.1 (10x Genomics, Cat# PN-1000121) according to the manufacturer’s protocol for 10000 cell recovery. Results: paraxial mesoderm, lateral plate mesoderm, and their derivatives were detected in bmp4 explants, and NMP, somite and neural cell fates were detected in bmp4 induced human embryoids. Conclusions: A well-organized caudal-like structure was induced in bmp4 explants and caudal cell fates were specified in BMP4 induced human embryoids. bmp4 injected zebrafish explants or BMP4 induced human embryoids were harvested for scRNA-seq.