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  • Preparation of Primary Astrocyte Culture Derived from Human Glioblastoma Multiforme Specimen.

Preparation of Primary Astrocyte Culture Derived from Human Glioblastoma Multiforme Specimen.

Bio-protocol (2017-04-20)
Mansoureh Hashemi, Mahmoudreza Hadjighassem
ABSTRACT

Glioblastoma multiforme (GBM) is a grade 4 astrocytoma tumor in central nervous system. Astrocytes can be isolated from human GBM. Study of astrocytes can provide insights about the formation, progression and recurrence of glioblastoma. For using isolated astrocytes, new studies can be designed in the fields of pharmacology, neuroscience and neurosurgery for glioblastoma treatment. This protocol describes the details for preparing high purity primary astrocytes from human GBM. Tumor tissue is disrupted using mechanical dissociation and chemical digestion in this protocol. 2 weeks after plating the cell suspension in culture, primary astrocytes are available for further subculturing and immunocytochemistry of S100-beta antigen.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Paraformaldehyde, reagent grade, crystalline
Sigma-Aldrich
Anti-S-100 antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Triton X-100, Molecular Biology
Sigma-Aldrich
Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture