Generation of Pulmonary Endothelial Progenitor Cells for Cell-based Therapy Using Interspecies Mouse–Rat Chimeras

In Vivo Generation of Bone Marrow from Embryonic Stem Cells in Interspecies Chimeras

“…Consistent with functional competency of ESC-derived bone marrow cells, transplantation of these cells into lethally-irradiated syngeneic mice prevented mortality and resulted in long-term contribution of ESC-derived cells to all hematopoietic cell lineages in the bone marrow and peripheral blood. Our results are consistent with recent studies demonstrating the ability of mouse ESCs to generate functional pancreatic, endothelial and kidney cells in interspecies mouse-rat chimeras (25, 27, 35). Interestingly, long-term contribution of donor BM cells to ST- HSCs and LT-HSCs of irradiated mice was high, supporting the ability of donor HSCs to self- renew.…”

“…C57BL/6 mice were purchased from Jackson Lab. Interspecies mouse-rat chimeras were generated using blastocyst complementation as described (35, 36). Briefly, blastocysts from SD rats were obtained at embryonic day 4.5 (E4.5), injected with fifteen GFP- labeled mouse ESC cells (ESC-GFP, C57BL/6 background) (32, 37) and transferred into pseudo pregnant SD rat females.…”

“…Read alignments, quality controls and false discovery rates were described previously (43, 44). Identification of cell clusters and quantification of cluster-specific gene expression in BM scRNAseq datasets was performed as described (1, 32, 35). To assess the transcriptomic similarity of ESC-derived cells from mouse- rat and mouse-mouse BM, the scRNAseq datasets were normalized with SCTransform and then integrated utilizing the canonical correlation analysis (CCA).…”

“…Using blastocyst complementation, mouse ESCs effectively produced hematopoietic cells in mice deficient for Kit or Flk1 (28, 34). ESC-derived endothelial progenitor cells from mouse-rat chimeras were indistinguishable from endogenous endothelial progenitor cells based on gene expression signatures and functional properties (35). While all these studies support the effectiveness of blastocyst complementation for differentiation of multiple cell types from ESCs/iPSCs in vivo , generation of functional bone marrow from ESCs in interspecies chimeras has not yet been achieved.…”

All Roads Lead to Rome? Resident or Interspecies Chimera-derived Pulmonary Endothelial Progenitors for Cell-based Therapy

“…In this issue of the Journal , Wang and colleagues (pp. 326–338 ) have identified additional heterogeneity within this cKIT + progenitor population consisting of Foxf1 (forkhead box F1)-positive or -negative cells by means of single-cell RNA-sequencing ( 12 ). FOXF1 + cKIT + cells, also present in the human lung, decrease in the adult and are enriched in genes related to angiogenesis and endothelial cell proliferation.…”