Immortalization and characterization of human myometrial cells from term-pregnant patients using a telomerase expression vector.
An examination of cellular processes involved in myometrial function has been greatly assisted by the use of human myometrial cells in primary culture. However, these cells can be used only for several passages before they senesce, and responses to various agents change with time in culture. The use of transformed cells is limited, as they can be polynucleated and can lose or gain chromosomes. We have developed three telomerase-immortalized cell lines from term-pregnant human myometrium to eliminate variability between passage numbers and allow genetic manipulations of myometrial cells to fully characterize signal pathways. These cells have a normal karyotype and were verified to be uterine smooth muscle by immunocytochemical staining for smooth muscle cell-specific alpha-actin and high affinity oxytocin antagonist binding sites. The three cell lines and the cells in primary culture from which they were derived were examined by cDNA microarray analysis. Of >10 000 expressed genes, there were consistent changes in the expression of approximately 1% in the three immortalized cell lines. We were unable to detect any significant differences between primary and immortalized cells in signal pathways such as epidermal growth factor-stimulated epidermal growth factor receptor phosphorylation, insulin-stimulated Akt phosphorylation, oxytocin and lysophosphatidic acid-stimulated extracellular signal-regulated kinase 1 and 2 phosphorylation, myosin light chain phosphorylation, and interleukin-1 induction of IkappaBalpha degradation. The immortalized cells should be useful for a range of studies, including high throughput analyses of the effects of environmental agents on the human myometrium.
Soloff
Melvyn S
MS
Department of Obstetrics and GynecologySealy Center for Molecular Science, University of Texas Medical Branch, Galveston, TX 77555-1062, USA. msoloff@utmb.edu
Jeng
Yow-Jiun
YJ
Ilies
Monica
M
Soloff
Solweig L
SL
Izban
Michael G
MG
Wood
Thomas G
TG
Panova
Neli I
NI
Velagaleti
Gopalrao V N
GV
Anderson
Garland D
GD
D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D005822:Genetic Vectors; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007375:Interleukin-1 / Q000494:pharmacology; D007621:Karyotyping; D008246:Lysophospholipids / Q000378:metabolism; D009215:Myometrium / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D018994:Myosin Light Chains / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D020411:Oligonucleotide Array Sequence Analysis; D010121:Oxytocin / Q000378:metabolism; D011247:Pregnancy; D015398:Signal Transduction / Q000502:physiology; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D007375:Interleukin-1; D008246:Lysophospholipids; D018994:Myosin Light Chains; C000606204:NFKBIA protein, human; D000072000:NF-KappaB Inhibitor alpha; D010121:Oxytocin; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D048049:Extracellular Signal-Regulated MAP Kinases; D019098:Telomerase; C032881:lysophosphatidic acid