Characterization of long-term cultured murine submandibular gland epithelial cells

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

研究成果: Article

4 引用 (Scopus)

抄録

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.

元の言語English
記事番号e0147407
ジャーナルPLoS One
11
発行部数1
DOI
出版物ステータスPublished - 2016 1 1

Fingerprint

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)

これを引用

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

:: PLoS One, 巻 11, 番号 1, e0147407, 01.01.2016.

研究成果: Article

@article{ce723debc8734457ba6ba769f9fa491c,
title = "Characterization of long-term cultured murine submandibular gland epithelial cells",
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.",
author = "Kazuhiro Ikeura and Tetsuya Kawakita and Kazuyuki Tsunoda and Taneaki Nakagawa and Kazuo Tsubota",
year = "2016",
month = "1",
day = "1",
doi = "10.1371/journal.pone.0147407",
language = "English",
volume = "11",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "1",

}

TY - JOUR

T1 - Characterization of long-term cultured murine submandibular gland epithelial cells

AU - Ikeura, Kazuhiro

AU - Kawakita, Tetsuya

AU - Tsunoda, Kazuyuki

AU - Nakagawa, Taneaki

AU - Tsubota, Kazuo

PY - 2016/1/1

Y1 - 2016/1/1

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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84958191038&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84958191038&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0147407

DO - 10.1371/journal.pone.0147407

M3 - Article

C2 - 26800086

AN - SCOPUS:84958191038

VL - 11

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 1

M1 - e0147407

ER -