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Relationship of the demonstration of intermediate filament protein to kinetics of three human neuroepithelial tumor cell lines. Lack of neural-related proteins in most cells in S phase: a double-labeled immunohistochemical study on matrix cultures.

Publication ,  Journal Article
Ibayashi, N; Herman, MM; Boyd, JC; Bigner, DD; Friedman, HS; Collins, VP; Donoso, LA; Rubinstein, LJ
Published in: Lab Invest
September 1989

The immunocytochemical demonstration of intermediate filament proteins in three human neuroepithelial tumor cell lines maintained in vitro on a three-dimensional matrix was correlated with the proportion of cells in S phase. The cell lines of a medulloblastoma (D283 Med), a retinoblastoma (WERI-Rb1), and an astrocytic glioma (U-251 MG) were cultured in an organ culture system, pulse-fed with bromodeoxyuridine, and double-labeled by immunoperoxidase and by the avidin-biotin peroxidase complex method for bromodeoxyuridine and for intermediate filament proteins [each triplet of neurofilament proteins, as well as vimentin and glial fibrillary acidic (GFA) protein] using eight different antibodies. The average percentages of bromodeoxyuridine-labeled cells for the D283 Med, WERI-Rb1, and U-251 MG lines were respectively, 25, 32, and 12% 30 minutes after pulse labeling. In the D283 Med line, 15- greater than 95% of the cells were positive for each neurofilament protein, and 80% of the cells were positive for vimentin; less than 10% of the cells in S phase were positive with each of the five antineurofilament protein monoclonal antibodies (Mabs), but 20% of the vimentin-positive cells were in S phase. In the WERI-Rb1 line, 44 and greater than 96% of the cells were positive for the high-molecular-weight neurofilament subunit and high- and middle-molecular-weight neurofilament subunits proteins, respectively, but only 5% of the high-molecular-weight neurofilament positive cells were in S phase. In the U-251 MG line, 37 and 98% of the cells were positive for GFA protein and vimentin, respectively; only 3% of the GFA protein-positive cells, but 13% of the vimentin-positive cells, were in S phase. The results indicate that, when maintained in a matrix culture system, most cells in S phase in these lines lack markers of neural differentiation.

Duke Scholars

Published In

Lab Invest

ISSN

0023-6837

Publication Date

September 1989

Volume

61

Issue

3

Start / End Page

310 / 318

Location

United States

Related Subject Headings

  • Vimentin
  • Tumor Cells, Cultured
  • Retinoblastoma
  • Pathology
  • Neurofilament Proteins
  • Medulloblastoma
  • Interphase
  • Intermediate Filament Proteins
  • Immunoenzyme Techniques
  • Humans
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Ibayashi, N., M. M. Herman, J. C. Boyd, D. D. Bigner, H. S. Friedman, V. P. Collins, L. A. Donoso, and L. J. Rubinstein. “Relationship of the demonstration of intermediate filament protein to kinetics of three human neuroepithelial tumor cell lines. Lack of neural-related proteins in most cells in S phase: a double-labeled immunohistochemical study on matrix cultures.Lab Invest 61, no. 3 (September 1989): 310–18.

Published In

Lab Invest

ISSN

0023-6837

Publication Date

September 1989

Volume

61

Issue

3

Start / End Page

310 / 318

Location

United States

Related Subject Headings

  • Vimentin
  • Tumor Cells, Cultured
  • Retinoblastoma
  • Pathology
  • Neurofilament Proteins
  • Medulloblastoma
  • Interphase
  • Intermediate Filament Proteins
  • Immunoenzyme Techniques
  • Humans