End-stage lung disease is the third leading cause of death in the United States. Lung transplantation is the only definitive treatment to replace lungs in end-stage patients. However, complications are frequent, and there is a severe shortage of organs. Stem cell-based therapy supplying exogenous stem/progenitor cells to help lung repair or ultimately replacing diseased lung tissue have become prime translational goals. One of the major challenges before stem cell-based therapies can be realized is generating sufficient quantities of the appropriate progenitor and mature cells. We have previously shown that three-dimensional branching lung organoids can be generated from human pluripotent stem cells (hPSCs, including embryonic stem cells and induced pluripotent stem cells). Here we show that we can isolate and grow cells from these organoids with a phenotype and expression pattern compatible with human distal lung bud tip progenitor cells (SOX2+SOX9+). These cells, which we call putative distal tip progenitors (pDTPs) could be maintained as a homogenous population, passaged for > 12 months, frozen and thawed. The cells maintained active proliferation and expanded > 90 fold in 2 weeks even at passage 38. In vitro expanded pDTPs attach to, flatten and survive for at least 7 days in de-epithelialized rat lungs ex vivo and for at least 2 weeks in injured immunodeficient mouse lung in vivo after delivery in the airway. Furthermore, they can be differentiated in vitro into mature basal cells (NGFR+P63+CD104+CD49f+), the stem cells of airway, and further differentiated into ciliated, goblet, and club cells. The in vitro expanded basal cells can engraft and repopulate de-epithelialized mouse tracheas ex vivo. The capacity to generate an unlimited number of distal lung bud tip progenitor cells and basal cells from hPSCs provides a potential source of cells for stem cell therapies and regenerative medicine in the lung.
Physiology 2019 (Aberdeen, UK) (2019) Proc Physiol Soc 43, SA039
Research Symposium: Unlimited expansion of putative distal lung bud tip progenitor cells from human pluripotent stem cells and their ex vivo and in vivo engraftment
Y. Chen1,2,3, H. Liu4,5, N. Dorrello6,7, K. Cunningham7, G. Vunjak-Novakovic7,4, H. Snoeck4,5,8,9
1. Medicine, University of Southern California, Los Angeles, California, United States. 2. Hastings Center for Pulmonary Research, University of Southern California, Los Angeles, California, United States. 3. Pulmonary, Critical Care and Sleep Medicine, University of Southern California, Los Angeles, California, United States. 4. Medicine, Columbia University Irving Medical Center, New York, New York, United States. 5. Columbia Center for Human Development, Columbia University Irving Medical Center, New York, New York, United States. 6. Pediatrics, Columbia University Irving Medical Center, New York, New York, United States. 7. Biomedical Engineering, Columbia University Irving Medical Center, New York, New York, United States. 8. Columbia Center for Translational Immunology, Columbia University Irving Medical Center, New York, New York, United States. 9. Microbiology and Immunology, Columbia University Irving Medical Center, New York, New York, United States.
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Where applicable, experiments conform with Society ethical requirements.