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 - 1993/5
Y1 - 1993/5
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. © 1993 Wiley‐Liss, Inc.
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. © 1993 Wiley‐Liss, Inc.
KW - Diffuse neuroendocrine system
KW - Immunoreactive calcitonin
KW - Small cell lung cancer
KW - general NE markers
KW - specific products
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U2 - 10.1002/ar.1092360128
DO - 10.1002/ar.1092360128
M3 - Article
C2 - 8389531
AN - SCOPUS:0027300109
VL - 236
SP - 231
EP - 240
JO - Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology
JF - Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology
SN - 0003-276X
IS - 1
ER -