Long-term selective culture of hamster pulmonary endocrine cells

R. I. Linnoila, A. F. Gazdar, K. Funa, K. L. Becker

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We describe, for the first time, the development of a technique for a long-term selective culture of endocrine (PE) cells from the lungs of normal animals. Epithelial cells were isolated from 1-day-old hamster lungs through mechanical and enzymatic dissociation with collagenase type II. Cells were then cultured in HITES medium which contained RPMI 1640, hydrocortisone, insulin, transferrin, estradiol, sodium selenite, and supplemented with 5% fetal bovine serum (FBS), or medium which contained HITES medium supplemented with bovine serum albumin, phosphoethanolamine, arginine vasopressin, bombesin, and 2% FBS (9N). HITES medium, originally developed for establishment and long-term culture of human small cell lung cancer (SCLC) cell lines, allowed propagation of normal hamster PE cells up to 12 months as a mixed floating-attached cell culture. No difference was noted in the results using HITES or 9N. By 3 months, 80% of the cultured cells contained characteristic dense-core (endocrine type) granules. The cultured PE cells also expressed creatine kinase brain isoenzyme, and general NE markers including neuron specific enolase, and amine handling enzyme activity within the range of SCLC cell lines. Moreover, cultured PE cells contained and secreted immunoreactive calcitonin (iCT) which had a molecular profile similar to that of intact hamster lung. This long-term culture technique should markedly assist in elucidating the role of PE cells in health and disease.

Original languageEnglish (US)
Pages (from-to)231-240
Number of pages10
JournalAnatomical Record
Volume236
Issue number1
StatePublished - 1993

Fingerprint

Endocrine Cells
hamsters
Cricetinae
serum
Cultured Cells
lungs
Small Cell Lung Carcinoma
Lung
cancer
BB Form Creatine Kinase
cultured cells
Sodium Selenite
health and disease
Cell Line
Bombesin
Culture Techniques
selenite
Phosphopyruvate Hydratase
Arginine Vasopressin
fetal bovine serum

Keywords

  • Diffuse neuroendocrine system
  • general NE markers
  • Immunoreactive calcitonin
  • Small cell lung cancer
  • specific products

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Anatomy

Cite this

Linnoila, R. I., Gazdar, A. F., Funa, K., & Becker, K. L. (1993). Long-term selective culture of hamster pulmonary endocrine cells. Anatomical Record, 236(1), 231-240.

Long-term selective culture of hamster pulmonary endocrine cells. / Linnoila, R. I.; Gazdar, A. F.; Funa, K.; Becker, K. L.

In: Anatomical Record, Vol. 236, No. 1, 1993, p. 231-240.

Research output: Contribution to journalArticle

Linnoila, RI, Gazdar, AF, Funa, K & Becker, KL 1993, 'Long-term selective culture of hamster pulmonary endocrine cells', Anatomical Record, vol. 236, no. 1, pp. 231-240.
Linnoila RI, Gazdar AF, Funa K, Becker KL. Long-term selective culture of hamster pulmonary endocrine cells. Anatomical Record. 1993;236(1):231-240.
Linnoila, R. I. ; Gazdar, A. F. ; Funa, K. ; Becker, K. L. / Long-term selective culture of hamster pulmonary endocrine cells. In: Anatomical Record. 1993 ; Vol. 236, No. 1. pp. 231-240.
@article{ec7949887584443996511d5f63ce200e,
title = "Long-term selective culture of hamster pulmonary endocrine cells",
abstract = "We describe, for the first time, the development of a technique for a long-term selective culture of endocrine (PE) cells from the lungs of normal animals. Epithelial cells were isolated from 1-day-old hamster lungs through mechanical and enzymatic dissociation with collagenase type II. Cells were then cultured in HITES medium which contained RPMI 1640, hydrocortisone, insulin, transferrin, estradiol, sodium selenite, and supplemented with 5{\%} fetal bovine serum (FBS), or medium which contained HITES medium supplemented with bovine serum albumin, phosphoethanolamine, arginine vasopressin, bombesin, and 2{\%} FBS (9N). HITES medium, originally developed for establishment and long-term culture of human small cell lung cancer (SCLC) cell lines, allowed propagation of normal hamster PE cells up to 12 months as a mixed floating-attached cell culture. No difference was noted in the results using HITES or 9N. By 3 months, 80{\%} of the cultured cells contained characteristic dense-core (endocrine type) granules. The cultured PE cells also expressed creatine kinase brain isoenzyme, and general NE markers including neuron specific enolase, and amine handling enzyme activity within the range of SCLC cell lines. Moreover, cultured PE cells contained and secreted immunoreactive calcitonin (iCT) which had a molecular profile similar to that of intact hamster lung. This long-term culture technique should markedly assist in elucidating the role of PE cells in health and disease.",
keywords = "Diffuse neuroendocrine system, general NE markers, Immunoreactive calcitonin, Small cell lung cancer, specific products",
author = "Linnoila, {R. I.} and Gazdar, {A. F.} and K. Funa and Becker, {K. L.}",
year = "1993",
language = "English (US)",
volume = "236",
pages = "231--240",
journal = "Anatomical Record",
issn = "0003-276X",
publisher = "John Wiley and Sons Inc.",
number = "1",

}

TY - JOUR

T1 - Long-term selective culture of hamster pulmonary endocrine cells

AU - Linnoila, R. I.

AU - Gazdar, A. F.

AU - Funa, K.

AU - Becker, K. L.

PY - 1993

Y1 - 1993

N2 - We describe, for the first time, the development of a technique for a long-term selective culture of endocrine (PE) cells from the lungs of normal animals. Epithelial cells were isolated from 1-day-old hamster lungs through mechanical and enzymatic dissociation with collagenase type II. Cells were then cultured in HITES medium which contained RPMI 1640, hydrocortisone, insulin, transferrin, estradiol, sodium selenite, and supplemented with 5% fetal bovine serum (FBS), or medium which contained HITES medium supplemented with bovine serum albumin, phosphoethanolamine, arginine vasopressin, bombesin, and 2% FBS (9N). HITES medium, originally developed for establishment and long-term culture of human small cell lung cancer (SCLC) cell lines, allowed propagation of normal hamster PE cells up to 12 months as a mixed floating-attached cell culture. No difference was noted in the results using HITES or 9N. By 3 months, 80% of the cultured cells contained characteristic dense-core (endocrine type) granules. The cultured PE cells also expressed creatine kinase brain isoenzyme, and general NE markers including neuron specific enolase, and amine handling enzyme activity within the range of SCLC cell lines. Moreover, cultured PE cells contained and secreted immunoreactive calcitonin (iCT) which had a molecular profile similar to that of intact hamster lung. This long-term culture technique should markedly assist in elucidating the role of PE cells in health and disease.

AB - We describe, for the first time, the development of a technique for a long-term selective culture of endocrine (PE) cells from the lungs of normal animals. Epithelial cells were isolated from 1-day-old hamster lungs through mechanical and enzymatic dissociation with collagenase type II. Cells were then cultured in HITES medium which contained RPMI 1640, hydrocortisone, insulin, transferrin, estradiol, sodium selenite, and supplemented with 5% fetal bovine serum (FBS), or medium which contained HITES medium supplemented with bovine serum albumin, phosphoethanolamine, arginine vasopressin, bombesin, and 2% FBS (9N). HITES medium, originally developed for establishment and long-term culture of human small cell lung cancer (SCLC) cell lines, allowed propagation of normal hamster PE cells up to 12 months as a mixed floating-attached cell culture. No difference was noted in the results using HITES or 9N. By 3 months, 80% of the cultured cells contained characteristic dense-core (endocrine type) granules. The cultured PE cells also expressed creatine kinase brain isoenzyme, and general NE markers including neuron specific enolase, and amine handling enzyme activity within the range of SCLC cell lines. Moreover, cultured PE cells contained and secreted immunoreactive calcitonin (iCT) which had a molecular profile similar to that of intact hamster lung. This long-term culture technique should markedly assist in elucidating the role of PE cells in health and disease.

KW - Diffuse neuroendocrine system

KW - general NE markers

KW - Immunoreactive calcitonin

KW - Small cell lung cancer

KW - specific products

UR - http://www.scopus.com/inward/record.url?scp=0027300109&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027300109&partnerID=8YFLogxK

M3 - Article

C2 - 8389531

AN - SCOPUS:0027300109

VL - 236

SP - 231

EP - 240

JO - Anatomical Record

JF - Anatomical Record

SN - 0003-276X

IS - 1

ER -