mESC-derived donor cells in culture and 15 weeks after transplantation. mESC-derived donor cells in culture and 15 weeks after transplantation

The goal of this experiment was to compare the transcriptome of transplanted mouse ESC-derived cells and endogenous epithelial cells at 15 weeks post transplantation into a syngeneic wild type recipient. In addition these cells were compared to ESC-derived and primary mouse tip-like cells grown in culture, to determine how cultured cells changed followeing transplantation. Overall design: A 9 week old wild type 129X1/S1 mouse was given 1.5U/kg bleomycin intratracheally. 10 days later 5e5 ESC-derived tip-like cells were transplanted intratracheally into the same mouse. At 15 weeks post-transplantation the lungs of this mouse were digested and sorted using FACS. We collected both endogenous and donor-derived epithelial cells from the same lung. To analyze pre-transplantation donor populations, we collected passage 4 ESC-derived and passage 3 primary tip-like cells grown in Lung progenitor media for 6 days since passaging. Finally, we collected D13 ESC-derived epithelial cells from the WBRF lung specification protocol as outlined in Herriges et al., 2022.

本实验旨在比较移植后15周、植入同基因野生型受体体内的小鼠胚胎干细胞（ESC）衍生细胞与内源性上皮细胞的转录组。此外，本研究还将上述细胞与体外培养的ESC衍生细胞及原代小鼠尖端样细胞进行对比，以明确移植后培养细胞的变化情况。总体实验设计：选取1只9周龄野生型129X1/S1小鼠，经气管内给予1.5U/kg博莱霉素。10天后，向同一只小鼠气管内移植5×10^5个ESC衍生的尖端样细胞。移植后15周，对该小鼠的肺脏进行消化处理，并通过荧光激活细胞分选术（FACS）进行分选。我们从同一肺脏中同时收集内源性上皮细胞与供体来源的上皮细胞。为分析移植前的供体细胞群，我们收集了在肺祖细胞培养基中传代后培养6天的第4代ESC衍生细胞及第3代原代尖端样细胞。最后，我们按照Herriges等人2022年发表的WBRF肺谱系特化方案，收集了第13天的ESC衍生上皮细胞。