Generation of SV40-transformed rabbit tracheal-epithelial-cell-derived blastocyst by somatic cell nuclear transfer

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...

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Bibliographic Details
Published in:Cell and tissue research 2012-02, Vol.347 (2), p.357-367
Main Authors: de Semir, D, Maurisse, R, Du, F, Xu, J, Yang, X, Illek, B, Gruenert, D. C
Format: Article
Language:English
Subjects:
Analysis
Animal models
Animals
Biomedical and Life Sciences
Biomedicine
blastocyst
Blastocyst - cytology
Blastocyst - metabolism
blastocysts
Cell Line, Transformed
Cells
Cells, Cultured
Chloride transport
Cyclic AMP - metabolism
Cystic fibrosis
Cystic Fibrosis - genetics
Cystic Fibrosis - metabolism
Cystic Fibrosis - therapy
Cystic fibrosis transmembrane conductance regulator
Cystic Fibrosis Transmembrane Conductance Regulator - genetics
Cystic Fibrosis Transmembrane Conductance Regulator - metabolism
Cytochemistry
Cytokeratin
Cytology
Epithelial cells
Epithelial Cells - metabolism
Epithelium
Gene expression
genes
Hereditary diseases
Human Genetics
ion transport
Membranes
Molecular Medicine
Nuclear Transfer Techniques
Proteins
Proteomics
Rabbits
Regular Article
Respiratory tract
Simian virus 40 - genetics
Simian virus 40 - metabolism
somatic cell nuclear transfer
somatic cells
Tight junctions
Trachea - metabolism
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Summary: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.
ISSN:0302-766X
1432-0878
DOI:10.1007/s00441-011-1296-1