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Hydrogel Environment Supports Cell Culture Expansion of a Grade IV Astrocytoma
- Source :
- Neurochemical Research. 42:2610-2624
- Publication Year :
- 2017
- Publisher :
- Springer Science and Business Media LLC, 2017.
-
Abstract
- Malignant astrocytomas are aggressive cancers of glial origin that can develop into invasive brain tumors. The disease has poor prognosis and high recurrence rate. Astrocytoma cell lines of human origin are an important tool in the experimental pathway from bench to bedside because they afford a convenient intermediate system for in vitro analysis of brain cancer pathogenesis and treatment options. We undertook the current study to determine whether hydrogel culture methods could be adapted to support the growth of astrocytoma cell lines, thereby facilitating a system that may be biologically more similar to in vivo tumor tissue. Our experimental protocols enabled maintenance of Grade IV astrocytoma cell lines in conventional monolayer culture and in the extracellular matrix hydrogel, Geltrex™. Light and fluorescence microscopy showed that hydrogel environments promoted cellular reorganization from dispersed cells into multilayered aggregates. Transmission electron microscopy revealed the prevalence of autophagy and nuclear membrane distortions in both culture systems. Analysis of microarray Gene Expression Omnibus (GEO) DataSets highlighted expression of genes implicated in pathways for cancer progression and autophagy. A pilot quantitative polymerase chain reaction (qPCR) analysis of the autophagic biomarkers, Beclin 1 (BECN1) and microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B), with two reference genes (beta actin, ACTB; glyceraldehyde 3-phosphate dehydrogenase, GAPDH), uncovered a relative increase of BECN1 and LC3B in hydrogel cultures of astrocytoma as compared to the monolayer. Taken together, results establish that ultrastructural and molecular characteristics of autophagy are features of this astrocytoma cell line, and that hydrogel culture systems can afford novel opportunities for in vitro studies of glioma.
- Subjects :
- 0301 basic medicine
Astrocytoma
Biology
Biochemistry
Hydrogel, Polyethylene Glycol Dimethacrylate
Article
Extracellular matrix
03 medical and health sciences
Cellular and Molecular Neuroscience
Cell Line, Tumor
Glioma
Tumor Microenvironment
medicine
Humans
Gene Regulatory Networks
Brain Neoplasms
Autophagy
General Medicine
BECN1
medicine.disease
Cell biology
030104 developmental biology
Real-time polymerase chain reaction
Cell culture
Neoplasm Grading
MAP1LC3B
Subjects
Details
- ISSN :
- 15736903 and 03643190
- Volume :
- 42
- Database :
- OpenAIRE
- Journal :
- Neurochemical Research
- Accession number :
- edsair.doi.dedup.....455aadcd009e3868424ff4a483899639