Generation of SV40-transformed rabbit tracheal-epithelial-cell-derived blastocyst by somatic cell nuclear transfer |
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| Authors: | D. de Semir R. Maurisse F. Du J. Xu X. Yang B. Illek D. C. Gruenert |
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| Affiliation: | (1) Department of Otolaryngology—Head and Neck Surgery, University of California, San Francisco, Mount Zion Cancer Center, 2340 Sutter Street, Box 1330, San Francisco, CA 94115, USA;(2) Department of Laboratory Medicine, Helen Diller Family Comprehensive Cancer Center, Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, Institute for Human Genetics, University of California, San Francisco, USA;(3) California Pacific Medical Center Research Institute, San Francisco, Calif., USA;(4) Evergen Biotechnologies, 1392 Storrs Road, Unit 4213 Storrs, Conn., USA;(5) Institute for Regenerative Medicine, University of Connecticut, Storrs, Conn., USA;(6) Children’s Hospital Oakland Research Institute, Oakland, Calif., USA;(7) Department of Pediatrics, University of Vermont College of Medicine, Burlington, Vt., USA;(8) Present address: Medicen, 6 Rue Alexandre Cabanel, 75015 Paris, France;(9) Present address: Renova Life, 387 Technology Drive, College Park, Md., USA; |
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| Abstract: | The prospect of developing large animal models for the study of inherited diseases, such as cystic fibrosis (CF), through somatic cell nuclear transfer (SCNT) has opened up new opportunities for enhancing our understanding of disease pathology and for identifying new therapies. Thus, the development of species-specific in vitro cell systems that will provide broader insight into organ- and cell-type-specific functions relevant to the pathology of the disease is crucial. Studies have been undertaken to establish transformed rabbit airway epithelial cell lines that display differentiated features characteristic of the primary airway epithelium. This study describes the successful establishment and characterization of two SV40-transformed rabbit tracheal epithelial cell lines. These cell lines, 5RTEo- and 9RTEo-, express the CF transmembrane conductance regulator (CFTR) gene, retain epithelial-specific differentiated morphology and show CFTR-based cAMP-dependent Cl− ion transport across the apical membrane of a confluent monolayer. Immunocytochemical analysis indicates the presence of airway cytokeratins and tight-junction proteins in the 9RTEo- cell line after multiple generations. However, the tight junctions appear to diminish in their efficacy in both cell lines after at least 100 generations. Initial SCNT studies with the 9RTEo- cells have revealed that SV40-transformed rabbit airway epithelial donor cells can be used to generate blastocysts. These cell systems provide valuable models for studying the developmental and metabolic modulation of CFTR gene expression and rabbit airway epithelial cell biology. |
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