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Immortal, developmentally arrested human fetal kidney cell lines created by retroviral expression of human papilloma
I A Drummond1, P Goodyer, V P Sukhatme
1Department of Medicine/Renal Division, Beth Israel Deaconess Medical Center/Harvard Medical School, Boston, MA 02215, USA.
Aims:
For studies of kidney organogenesis, clonal human fetal kidney (HFK) cell lines that represent various stages of kidney epithelial development would provide valuable tools for in vitro approaches to kidney cell differentiation.
Methods:
Cell cultures from the cortices of 12- to 16-week HFKs were immortalized by retroviral expression of human papilloma virus (HPV) E6/E7 genes, and clonal isolates were characterized with regard to morphology, expression of molecular markers, growth on different matrices, and their ability to differentiate further in vitro.
Results:
Three immortal, clonal cell lines displayed homogeneous but distinct morphologies in vitro, ranging from mesenchymal to epithelial. Northern blotting with stage-specific markers allowed us to assign particular cell lines to discrete stages in kidney cell differentiation. These results were confirmed by assessing cell growth on type-I collagen versus Matrigel matrices. We have also demonstrated that for one of the cell lines fetal calf serum contains a differentiation factor that induced morphological differentiation and enhanced expression of epithelial-specific genes.
Conclusion:
Immortalization by HPV-E6/E7 expression preserved important phenotypic characteristics of fetal kidney cells. The creation of these lines provides useful tools for in vitro approaches to kidney growth and development.