Characterization of long-term cultured murine submandibular gland epithelial cells

Kazuhiro Ikeura, Tetsuya Kawakita, Kazuyuki Tsunoda, Taneaki Nakagawa, Kazuo Tsubota

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Purpose Human and rat salivary gland cell lines derived from tumors or genetic modification are currently available for research. Here, we attempted to culture and characterize longterm cultured cells spontaneously derived from wild type murine submandibular glands (SGs). Methods SGs were removed from 3-week-old C57B/6J female mice and dissociated by collagenase type 1 and hyaluronidase digestion. Isolated SG epithelial cells were cultured in low calcium, serum-free growth media in the presence of cholera toxin (CT) during early passages. Single-cell colonies were isolated by limiting dilution culture after 25 passages. Early- and late-stage cell cultures were characterized for keratin 14, keratin 18, α-smooth muscle actin, and p63 by immunostaining and quantitative real-time PCR analysis. Results SG epithelial cells cultured in optimized media maintained their proliferative ability and morphology for over 80 passages. Long-term cultured cells expressed keratin 14, keratin 18, and p63, indicative of an epithelial phenotype. Conclusions Epithelial cells originating from wild type murine SGs could be cultured for longer periods of time and remain phenotypically similar to ductal basal epithelium.

Original languageEnglish
Article numbere0147407
JournalPLoS One
Volume11
Issue number1
DOIs
Publication statusPublished - 2016 Jan 1

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keratin
Submandibular Gland
Keratin-14
Keratin-18
epithelial cells
Epithelial Cells
Cells
Cell culture
mice
cultured cells
Hyaluronoglucosaminidase
Cholera Toxin
Cultured Cells
hyaluronoglucosaminidase
Dilution
cholera toxin
collagenase
Muscle
Rats
Actins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Characterization of long-term cultured murine submandibular gland epithelial cells. / Ikeura, Kazuhiro; Kawakita, Tetsuya; Tsunoda, Kazuyuki; Nakagawa, Taneaki; Tsubota, Kazuo.

In: PLoS One, Vol. 11, No. 1, e0147407, 01.01.2016.

Research output: Contribution to journalArticle

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AU - Ikeura, Kazuhiro

AU - Kawakita, Tetsuya

AU - Tsunoda, Kazuyuki

AU - Nakagawa, Taneaki

AU - Tsubota, Kazuo

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N2 - Purpose Human and rat salivary gland cell lines derived from tumors or genetic modification are currently available for research. Here, we attempted to culture and characterize longterm cultured cells spontaneously derived from wild type murine submandibular glands (SGs). Methods SGs were removed from 3-week-old C57B/6J female mice and dissociated by collagenase type 1 and hyaluronidase digestion. Isolated SG epithelial cells were cultured in low calcium, serum-free growth media in the presence of cholera toxin (CT) during early passages. Single-cell colonies were isolated by limiting dilution culture after 25 passages. Early- and late-stage cell cultures were characterized for keratin 14, keratin 18, α-smooth muscle actin, and p63 by immunostaining and quantitative real-time PCR analysis. Results SG epithelial cells cultured in optimized media maintained their proliferative ability and morphology for over 80 passages. Long-term cultured cells expressed keratin 14, keratin 18, and p63, indicative of an epithelial phenotype. Conclusions Epithelial cells originating from wild type murine SGs could be cultured for longer periods of time and remain phenotypically similar to ductal basal epithelium.

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