Characterization of adherent primary cell lines from fresh human glioblastoma tissue, defining glial fibrillary acidic protein as a reliable marker in establishment of glioblastoma cell culture.
GFAP
glioblastoma
in vitro
primary cell line
Journal
Cancer reports (Hoboken, N.J.)
ISSN: 2573-8348
Titre abrégé: Cancer Rep (Hoboken)
Pays: United States
ID NLM: 101747728
Informations de publication
Date de publication:
04 2021
04 2021
Historique:
revised:
06
11
2020
received:
12
06
2020
accepted:
09
11
2020
pubmed:
1
12
2020
medline:
30
11
2021
entrez:
30
11
2020
Statut:
ppublish
Résumé
Primary adherent glioblastoma cell lines are an important tool in investigating cellular and molecular tumor biology, as well as treatment options for patients. The phenotypical and immunocytochemical characterization of primary cell lines from glioblastoma specimens during establishment is of great importance, in order to reliably identify these cell lines as primary glioblastoma cell lines. Sixteen primary adherent cell lines out of 34 glioblastoma samples (47%) were established and further characterized. For phenotypical characterization, morphology and growth characteristics of the cells were classified. The cell lines had a high growth rate with a doubling time of 2 to 14 days. Morphologically, the cells displayed spindle-form or polygonal to amorphous shapes and grow as monolayer or in foci without evidence of contact inhibition. The cells were able to migrate and to form colonies. For further characterization, the protein expression of the astrocyte-specific protein glial fibrillary acidic protein (GFAP), the glial marker S100B, the neuronal marker TUBB3, and malignancy marker VIM, as well as the progenitor markers NES and SOX2, the proliferation marker MKI67, and the fibroblast marker TE7 were determined. Based on the immunocytochemical validation criterion of a coexpression of GFAP and S100B, 15 out of these 16 cell lines (94%) were defined as primary glioblastoma cell lines (pGCL). All 15 pGCL expressed TUBB3 and VIM. NES and SOX2 were stained positively in 13/15 and 6/15 pGCL. MKI67 was expressed in 11/15 and TE7 in 2/15 pGCL. These results point out that in self-established primary adherent glioblastoma cell lines, the expression of the specific astrocytic and glial markers GFAP and S100B and of the malignancy and progenitor markers VIM, NES, and SOX2 has to be validated. These data show that primary cell lines of glioblastoma origin with high malignant potential are reliably to establish using standardized validation criteria.
Sections du résumé
BACKGROUND
Primary adherent glioblastoma cell lines are an important tool in investigating cellular and molecular tumor biology, as well as treatment options for patients.
AIM
The phenotypical and immunocytochemical characterization of primary cell lines from glioblastoma specimens during establishment is of great importance, in order to reliably identify these cell lines as primary glioblastoma cell lines.
METHODS AND RESULTS
Sixteen primary adherent cell lines out of 34 glioblastoma samples (47%) were established and further characterized. For phenotypical characterization, morphology and growth characteristics of the cells were classified. The cell lines had a high growth rate with a doubling time of 2 to 14 days. Morphologically, the cells displayed spindle-form or polygonal to amorphous shapes and grow as monolayer or in foci without evidence of contact inhibition. The cells were able to migrate and to form colonies. For further characterization, the protein expression of the astrocyte-specific protein glial fibrillary acidic protein (GFAP), the glial marker S100B, the neuronal marker TUBB3, and malignancy marker VIM, as well as the progenitor markers NES and SOX2, the proliferation marker MKI67, and the fibroblast marker TE7 were determined. Based on the immunocytochemical validation criterion of a coexpression of GFAP and S100B, 15 out of these 16 cell lines (94%) were defined as primary glioblastoma cell lines (pGCL). All 15 pGCL expressed TUBB3 and VIM. NES and SOX2 were stained positively in 13/15 and 6/15 pGCL. MKI67 was expressed in 11/15 and TE7 in 2/15 pGCL.
CONCLUSION
These results point out that in self-established primary adherent glioblastoma cell lines, the expression of the specific astrocytic and glial markers GFAP and S100B and of the malignancy and progenitor markers VIM, NES, and SOX2 has to be validated. These data show that primary cell lines of glioblastoma origin with high malignant potential are reliably to establish using standardized validation criteria.
Identifiants
pubmed: 33251771
doi: 10.1002/cnr2.1324
pmc: PMC8451382
doi:
Substances chimiques
Biomarkers, Tumor
0
GFAP protein, human
0
Glial Fibrillary Acidic Protein
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1324Informations de copyright
© 2020 The Authors. Cancer Reports published by Wiley Periodicals LLC.
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