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Immortalization of human small airway epithelial cells with characteristics of bronchioalveolar stem cells
Author(s) -
Gao Boning,
Huang Chunxian,
Sullivan James P,
spinola Monica,
Raso Maria Gabriela,
Wistuba Ignacio,
Shay Jerry,
Minna John D
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.lb340
Subject(s) - progenitor cell , stem cell , carcinogenesis , biology , lung , microbiology and biotechnology , matrigel , immunology , pathology , cancer research , angiogenesis , medicine , genetics , gene
Most of our current understanding of stem cells in the adult lung is based on mouse models while information in the human lung remains poorly defined. A major advance would be the ability to reproducibly grow human lung stem or progenitor cells to allow their systematic study and genetic manipulation. Toward this goal, we have worked out conditions to routinely immortalized (NB = 9) human small airway epithelial cells (HSAECs) from peripheral lung using Cdk4 and hTERT expression vectors in these cells. As an example strain HSAEC1 grows in serum free SAGM medium (Clonetics), forms colonies in liquid media efficiently, but does not form colonies in soft agar or tumors in SCID/NOD mice. HSAEC1 expresses lung basal/stem cell markers (aldehyde dehydrogenase, p63, Cytokeratin14 and 17, and is resistant to 1 mM naphthalene) but very low or undetectable Clara cell, Type1 and Type2 cell markers in SAGM, which are then expressed when grown in Matrigel culture or in serum. Finally, the cells can be genetically manipulated toward oncogenesis (with KRASV12, and or missense p53). We conclude, that we have prepared multiple, immortalized strains of human peripheral lung epithelial cells with many characteristics of lung stem or progenitor cells that are experimentally tractable for studying lung differentiation and tumorigenesis. This work is supported by NCI SPORE P50CA70907, and NASA/DOE NSCOR NNJ05HD36G, DE‐AI02‐05ER64068