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

Am J Respir Crit Care Med. 2021 Aug 1;204(3):326-338. doi: 10.1164/rccm.202003-0758OC.


Rationale: Although pulmonary endothelial progenitor cells (EPCs) hold promise for cell-based therapies for neonatal pulmonary disorders, whether EPCs can be derived from pluripotent embryonic stem cells (ESCs) or induced pluripotent stem cells remains unknown.Objectives: To investigate the heterogeneity of pulmonary EPCs and derive functional EPCs from pluripotent ESCs.Methods: Single-cell RNA sequencing of neonatal human and mouse lung was used to identify the heterogeneity of pulmonary EPCs. CRISPR/Cas9 gene editing was used to genetically label and purify mouse pulmonary EPCs. Functional properties of the EPCs were assessed after cell transplantation into neonatal mice with S52F Foxf1 mutation, a mouse model of alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV). Interspecies mouse-rat chimeras were produced through blastocyst complementation to generate EPCs from pluripotent ESCs for cell therapy in ACDMPV mice.Measurements and Main Results: We identified a unique population of EPCs, FOXF1+cKIT+ EPCs, as a subset of recently described general capillary cells (gCAPs) expressing SMAD7, ZBTB20, NFIA, and DLL4 but lacking mature arterial, venous, and lymphatic markers. FOXF1+cKIT+ gCAPs are reduced in ACDMPV, and their transcriptomic signature is conserved in mouse and human lungs. After cell transplantation into the neonatal circulation of ACDMPV mice, FOXF1+cKIT+ gCAPs engraft into the pulmonary vasculature, stimulate angiogenesis, improve oxygenation, and prevent alveolar simplification. FOXF1+cKIT+ gCAPs, produced from ESCs in interspecies chimeras, are fully competent to stimulate neonatal lung angiogenesis and alveolarization in ACDMPV mice.Conclusions: Cell-based therapy using donor or ESC/induced pluripotent stem cell-derived FOXF1+cKIT+ endothelial progenitors may be considered for treatment of human ACDMPV.

Keywords: alveolar capillary dysplasia; blastocyst complementation; cell therapy; endothelial progenitor cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Animals, Newborn
  • CRISPR-Cas Systems
  • Chimera
  • Disease Models, Animal
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Endothelial Progenitor Cells / cytology*
  • Endothelial Progenitor Cells / metabolism
  • Endothelial Progenitor Cells / transplantation
  • Forkhead Transcription Factors / genetics
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Infant, Newborn
  • Mice
  • Persistent Fetal Circulation Syndrome / metabolism
  • Persistent Fetal Circulation Syndrome / pathology
  • Persistent Fetal Circulation Syndrome / therapy*
  • Pluripotent Stem Cells
  • RNA-Seq
  • Rats
  • Single-Cell Analysis
  • Stem Cell Transplantation*


  • Forkhead Transcription Factors
  • Foxf1 protein, mouse