Juxtacrine stimulation of normal and malignant human bladder epithelial cell proliferation

Ursula K. Ehmann, Martha Kennedy Terris

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Purpose: We developed a method for culturing normal and malignant human bladder epithelial cells for many generations. Materials and Methods: Cells from bladder washes or surgical specimens were plated in culture with lethally irradiated cells of the LA7 rat mammary tumor line (American Type Culture Collection, Bethesda, Maryland) at a confluent density and fed regularly. After cells from surgical specimens became confluent they were diluted and subcultured with fresh irradiated LA7 cells. The growth rate was measured by cell counts and cell sorter analysis. Expression of intermediate filaments was determined by immunocytochemical testing. Results: All 5 normal and 3 of the 4 tumor specimens developed into long-term cell strains. A single normal strain was carried through passage 11, amounting to 37 cell doublings. Cell numbers doubled in 2 days in medium with 0.5% serum and in 5.6 days in 5% serum. Plating efficiency was almost 100% and cloning efficiency was approximately 9%. LA7 conditioned medium did not stimulate bladder cell proliferation. Two tumor strains were carried through passage 9, amounting to 20 and 27 doublings, respectively. No cell strains expressed signs of senescence at culture termination. Normal and tumor strains expressed keratins 7, 10, 11, 18 and 19, and ZO1 tight junction protein but not vimentin or keratin 14. Umbrella cells comprised the uppermost cell layer in cultures from normal bladder. Conclusions: LA7 feeder cells stimulate human bladder cell proliferation for many generations in culture by a juxtacrine mechanism and promote the expression of differentiated traits.

Original languageEnglish (US)
Pages (from-to)735-741
Number of pages7
JournalJournal of Urology
Volume167
Issue number2 I
StatePublished - Jan 1 2002
Externally publishedYes

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Urinary Bladder
Epithelial Cells
Cell Proliferation
Keratin-10
Cell Count
Keratin-14
Keratin-7
Tight Junction Proteins
Feeder Cells
Neoplasms
Intermediate Filaments
Vimentin
Conditioned Culture Medium
Serum
Organism Cloning
Breast Neoplasms
Growth

Keywords

  • Bladder
  • Cell culture
  • Cell line
  • Epithelium
  • Keratin

ASJC Scopus subject areas

  • Urology

Cite this

Juxtacrine stimulation of normal and malignant human bladder epithelial cell proliferation. / Ehmann, Ursula K.; Terris, Martha Kennedy.

In: Journal of Urology, Vol. 167, No. 2 I, 01.01.2002, p. 735-741.

Research output: Contribution to journalArticle

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abstract = "Purpose: We developed a method for culturing normal and malignant human bladder epithelial cells for many generations. Materials and Methods: Cells from bladder washes or surgical specimens were plated in culture with lethally irradiated cells of the LA7 rat mammary tumor line (American Type Culture Collection, Bethesda, Maryland) at a confluent density and fed regularly. After cells from surgical specimens became confluent they were diluted and subcultured with fresh irradiated LA7 cells. The growth rate was measured by cell counts and cell sorter analysis. Expression of intermediate filaments was determined by immunocytochemical testing. Results: All 5 normal and 3 of the 4 tumor specimens developed into long-term cell strains. A single normal strain was carried through passage 11, amounting to 37 cell doublings. Cell numbers doubled in 2 days in medium with 0.5{\%} serum and in 5.6 days in 5{\%} serum. Plating efficiency was almost 100{\%} and cloning efficiency was approximately 9{\%}. LA7 conditioned medium did not stimulate bladder cell proliferation. Two tumor strains were carried through passage 9, amounting to 20 and 27 doublings, respectively. No cell strains expressed signs of senescence at culture termination. Normal and tumor strains expressed keratins 7, 10, 11, 18 and 19, and ZO1 tight junction protein but not vimentin or keratin 14. Umbrella cells comprised the uppermost cell layer in cultures from normal bladder. Conclusions: LA7 feeder cells stimulate human bladder cell proliferation for many generations in culture by a juxtacrine mechanism and promote the expression of differentiated traits.",
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