Generation of a human embryonic stem cell line encoding the cystic fibrosis mutation ΔF508, using preimplantation genetic diagnosis

Abstract

Human embryonic stem (hES) cells are pluripotent cells isolated from early human embryos. They can be grown in vitro and made to differentiate into many different cell types. These properties have suggested that they may be useful in cell replacement therapy for many degenerative diseases. However, if hES cells could also be manufactured with mutations significant in human disease, they could provide a powerful in-vitro tool for modelling disease processes and progression in a number of different cell types, as well as providing an ideal system for studying in-vitro toxicity and efficacy of drugs and other therapeutic systems such as gene therapy. Embryos with such mutations are generated as part of routine genetic testing during preimplantation genetic diagnosis, providing the opportunity to generate cell lines with significant mutations. A human embryonic stem cell line homozygous for the most common mutation leading to cystic fibrosis in humans (delta F508) has been generated and characterized. This cell line has the same morphology and expresses proteins typical of other unaffected hES cell lines. This cell line represents an important in-vitro tool for understanding the pathophysiology of cystic fibrosis, and presents exciting opportunities to test the efficacy and toxicity of new therapies relevant to CF.

Stem cells and cell therapies for cystic fibrosis and other lung diseases

“…Comparable studies with human ES cells are limited in the United States due to current restrictions on human ES cell use. A human deltaF 508 embryonic stem cell line has been established in England [38]. These cells exhibit normal morphology and protein expression compared with other ES cell lines but have not been studied in detail.…”

Pluripotent Stem Cells in Research and Treatment of Hemoglobinopathies

“…Disease-specific human ESCs have been generated from embryos with genetic mutations causing diseases like thalassemia, Fanconi anemia, and cystic fibrosis (Pickering et al 2005;Verlinsky et al 2005;Mateizel et al 2006;Ben-Yosef et al 2008). However, affected embryos diagnosed through preimplantation testing are the prerequisite for deriving disease-relevant human ESCs, and because only a subset of predominantly Mendelian genetic disorders are subject to such testing, it is not feasible to obtain human ESCs for any specific disease of interest.…”

Mesenchymal Stem Cells: Possibilities of New Treatment Options

“…The hESC lines with different DNA content (normal, carrier, affected, and different polymorphic combinations) will be needed in the future for research and clinical use with regenerative purposes. PGD is a valuable source of embryos to make hESC lines both diseased and normal by using discarded and surplus embryos, respectively (Galat et al 2004 ;Kukharenko et al 2004 ;Pickering et al 2005 ;Verlinsky et al 2005 ; Mateizel et al 2006 ;Eiges et al 2007 ;Peura et al 2008 ;Tropel et al 2010 ;Frumkin et al 2010 ;Bradley et al 2011 ) . PGD-tested embryos can be an alternative source for normal euploid lines as well, possibly due to mosaic embryos having aneuploid and euploid cells or in vitro selection in favor of euploid cells (Lavon et al 2008 ;Taei et al 2010 ) .…”