Abstract
The HEK 293 cell line (293 cells) was derived from human embryonic kidney (HEK) cells grown in tissue culture. 293 cells are very easy to grow and transfect and have been widely used in cell biological research for many years. 293 cells have many of the properties of immature neurons, suggesting that they represent a transformed neuronal cell present in the original kidney culture, and they are not useful as an in vitro model for kidney cell function. The 293T cell line contains the SV(4)0 large T-antigen, which allows the episomal replication of transfected plasmids containing the SV40 origin of replication, and 293FT cells are a fast-growing variant. A recent report showed that introducing a set of transcription factors associated with pluripotency into human somatic cells can directly reprogr am them to produce induced pluripotent stem (iPS) cells. To date, however, iPS cells have not been generated from immortalized cells. We examined whether iPS cells could be generated from 293 FT cells transfected with four transcription factors (OCT4, SOX2, NANOG, and LIN28). The obtained cells morphologically resembled human ES cells, and showed a similar marker gene expression pattern. These cells had an impaired ability to differentiate, and formed immature ectodermal tumors after they were transplanted into nude mice. Thus, we could not derive fully reprogrammed iPS cells from 293FT cells. We conclude that the 293FT cells transduced with OCT4, SOX2, NANOG, and LIN28 produced aberrant ES-like cells.
Highlights
The HEK 293 cell line (293 cells) was derived from human embryonic kidney (HEK) cells grown in tissue culture. 293 cells are very easy to grow and transfect and have been widely used in cell biological research for many years. 293 cells have many of the properties of immature neurons, suggesting that they represent a transformed neuronal cell present in the original kidney culture, and they are not useful as an in vitro model for kidney cell function
The open reading frames of human OCT4, Some recent reports showed that introducing SOX2, NANOG, and LIN28 were amplified by a set of transcription factors associated with RT-PCR and subcloned, individually, into pluripotency into human somatic cells can pLenti 6.3/V5-TOPO (Invitrogen)
The RT-PCR was performed using the SuperScriptTMIII One-Step RT-PCR System with PlatinumR Taq DNA Polymerase (Invitrogen) for human ES cell markers, at 55°C for 30 min for the Results: ES cell-like colonies were generated from 293FT cells transduced with OCT4, SOX2, NANOG, and LIN28
Summary
The HEK 293 cell line (293 cells) was derived from human embryonic kidney (HEK) cells grown in tissue culture. 293 cells are very easy to grow and transfect and have been widely used in cell biological research for many years. 293 cells have many of the properties of immature neurons, suggesting that they represent a transformed neuronal cell present in the original kidney culture, and they are not useful as an in vitro model for kidney cell function. Colonies with a human ES cell-like morphology (iPS cell colonies) were picked 21-25 days after transduction. The RT-PCR was performed using the SuperScriptTMIII One-Step RT-PCR System with PlatinumR Taq DNA Polymerase (Invitrogen) for human ES cell markers, at 55°C for 30 min for the Results: ES cell-like colonies were generated from 293FT cells transduced with OCT4, SOX2, NANOG, and LIN28
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