Generation of Human Embryonic Kidney Cells with Extended In Vitro Life Span through Viral Oncogene Transfection

Abcouwer, Steve F.; Robinson, Polly S.; Goochee, Charles F.; Crow, Michael T.

doi:10.1038/nbt0989-939

Cited by 10 publications

(6 citation statements)

References 54 publications

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“…The human kidney cell lines ST-li, STt-4i, and 293 are all epithelial in origin (Abcouwer et al, 1989) but display no evidence of dome formation under the conditions examined in this study. This probably relates to their embryonic origin.…”

Section: Sample Collection and Assay Of Enzyme Activitiesmentioning

confidence: 93%

“…Cells ST-11 and STt-4i cells (Abcouwer et al, 1989) were analyzed at approximately 150 population doublings after crisis. A population of normal human embryonic kidney cells was obtained from NASA Space Bioprocessing Division, Johnson Space Center (Houston, TX).…”

Section: Methodsmentioning

confidence: 99%

“…Of note, is the observation in this study of different responses of OK and LLC-PK1 cells (both of proximal tubule origin) to the same differentiation inducin agents. in i efinite replicative potential (Abcouwer et al, 1989; In t I: is study, we investigated kidney-specific enzyme One potential reason for the f ifferent responses to inducing agents could be species specificity. A second potential reason is differences in the stage of kidney development when the cells were isolated.…”

Section: Sample Collection and Assay Of Enzyme Activitiesmentioning

confidence: 99%

“…The 293 cell line was generated by transfection of HEK cells with adenovirus DNA (Graham et al, 1977). The ST-11 cell line was generated by transfection of HEK cells with a plasmid encoding SV40 large T antigen, whereas generation of the STt-4i cell line involved transfection with a plasmid encoding both SV40 large T antigen and SV40 small T antigen (Abcouwer et al, 1989).…”

Section: Sample Collection and Assay Of Enzyme Activitiesmentioning

confidence: 99%

“…Several human kidney cell lines that could potentially serve as models for examining kidney cell function and kidneyspecific enzyme expression have been generated through oncogene transfection of normal human kidney cells. These include 293 cells (Graham et al, 1977), a cell line derived by transfection of adenovirus DNA into human embryonic kidney (HEK) cells, and the cell lines derived SV40 viral oncogenes into HEK cells (Abcouwer et al, 1989). There has been no previous examination of kidney-specific enzyme expression or kidney cell function for these cell cell lines.…”

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Kidney‐specific enzyme expression by human kidney cell lines generated through oncogene transfection

Robinson

Goochee

1991

Journal Cellular Physiology

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The human kidney cell line 293 was generated by transfection of adenovirus DNA into normal human embryonic kidney (HEK) cells (Graham et al., 1977), whereas the human kidney cell lines ST-1i and STt-4i were generated by transfection of HEK cells with plasmids encoding SV40 viral oncogenes (Abcouwer et al., 1989). In this study, we examined kidney-specific enzyme activity levels in 293, ST-1i, and STt-4i cells to determine their ability to exhibit kidney-specific gene expression. Enzymes examined were leucine aminopeptidase (LAP), gamma-glutamyl transpeptidase (gamma-GTP), and the disaccharidases trehalase and maltase. Enzymatic activity levels were compared to three other kidney cell lines (MDCK, OK, and LLC-PK1) as well as to normal human embryonic kidney (HEK) cells and the human hepatoma cell line, Hep G2. Modulation of kidney-specific enzyme activities was assessed in response to several differentiation-inducing agents (adenosine, n-butyric acid, hexamethylene bisacetamide (HMBA), dimethyl sulfoxide (DMSO), N,N'-dimethylformamide (DMF), isobutyl methyl xanthine (IBMX), di butyryl cAMP, and retinoic acid). ST-1i and STt-4i exhibit elevated levels of LAP, gamma-GTP, trehalase, and maltase, consistent with their kidney cell origin, whereas 293 cells exhibit elevated levels of just gamma-GTP and maltase. Maltase and gamma-GTP enzyme activities in ST-1i and STt-4i cells were very responsive to the various inducing agents; 293 cells were less responsive at the inducer concentrations examined. None of the three human cell lines formed domes under any of the experimental conditions. In summary, ST-1i and STt-4i are comparable to normal HEK cells in expression of kidney-specific enzymes and in responsiveness to differentiation-inducing agents, in spite of continued expression of SV40 oncogenes.

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Section: Sample Collection and Assay Of Enzyme Activitiesmentioning

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Section: Sample Collection and Assay Of Enzyme Activitiesmentioning

confidence: 99%

Section: Sample Collection and Assay Of Enzyme Activitiesmentioning

confidence: 99%

mentioning

confidence: 99%

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