Establishment of immortal swine kidney epithelial cells

Sungwook Kwak, Ji Eun Jung, Xun Jin, Sun Myung Kim, Tae Kyung Kim, Joong Seob Lee, Soo Yeon Lee, Xumin Pian, Seungkwon You, Hyunggee Kim, Yun Jaie Choi

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4 Citations (Scopus)

Abstract

Using normal swine kidney epithelial (SKE) cells that were shown to he senescent at passages 12 to 14, we have established one lifespan-extended cell line and two lifespan-extended cell lines by exogenous introduction of the human catalytic subunit of telomerase (hTERT) and simian virus 40 large T-antigen (SV40LT), all of which maintain epithelial morphology and express cytokeratin, a marker of epithelial cells. SV40LT- and hTERT-transduced immortal cell lines appeared to be smaller and exhibited more uniform morphology relative to primary and spontaneously immortalized SKE cells. We determined the in vitro lifespan of primary SKE cells using a standard 3T6 protocol. There were two steps of the proliferation harrier at 12 and 20, in which a majority of primary SKE cells appeared enlarged, flattened, vacuolated, and β-galactosidase-positive, all phenotypical characteristics of senescent cells. Lifespan-extended SKE cells were eventually established from most of the cellular foci, which is indicative of spontaneous cellular conversion at passage 23. Beyond passage 25, the rate of population doubling of the established cells gradually increased. At passage 30, immortal cell lines grew faster than primary counterpart cells in 10% FBS-DMEM culture conditions, and only SV40LT-transduced immortal cells grew faster than primary and other SKE immortal cells in 0.5% FBS-DMEM. These lifespan-extended SKE cell lines failed to grow in an anchorage-independent manner in soft-agar dishes. Hence, three immortalized swine kidney epithelial cells that are not transformed would be valuable biological tools for virus propagation and basic kidney epithelial cell research.

Original languageEnglish
Pages (from-to)51-58
Number of pages8
JournalAnimal Biotechnology
Volume17
Issue number1
DOIs
Publication statusPublished - 2006 May 1

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Keywords

  • Epithelial cell
  • Immortalization
  • Kidney
  • Swine
  • Telomerase. p53

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Biotechnology
  • Bioengineering

Cite this

Kwak, S., Jung, J. E., Jin, X., Kim, S. M., Kim, T. K., Lee, J. S., Lee, S. Y., Pian, X., You, S., Kim, H., & Choi, Y. J. (2006). Establishment of immortal swine kidney epithelial cells. Animal Biotechnology, 17(1), 51-58. https://doi.org/10.1080/10495390500461070