Establishment and Preliminary Characterization of Three Astrocytic Cells Lines Obtained from Primary Rat Astrocytes by Sub-Cloning.

Fabio Caradonna,Vincenzo Vitale,Pietro D’Oca,Carlo Maria Di Liegro,Tiziana Ferrara,Gabriella Schiera,Italia Di Liegro,Ilenia Cruciata,Riccardo Mormino,Simona Maria Angela Rizzo

doi:10.3390/genes11121502

Abstract

Gliomas are complex and heterogeneous tumors that originate from the glial cells of the brain. The malignant cells undergo deep modifications of their metabolism, and acquire the capacity to invade the brain parenchyma and to induce epigenetic modifications in the other brain cell types. In spite of the efforts made to define the pathology at the molecular level, and to set novel approaches to reach the infiltrating cells, gliomas are still fatal. In order to gain a better knowledge of the cellular events that accompany astrocyte transformation, we developed three increasingly transformed astrocyte cell lines, starting from primary rat cortical astrocytes, and analyzed them at the cytogenetic and epigenetic level. In parallel, we also studied the expression of the differentiation-related H1.0 linker histone variant to evaluate its possible modification in relation with transformation. We found that the most modified astrocytes (A-FC6) have epigenetic and chromosomal alterations typical of cancer, and that the other two clones (A-GS1 and A-VV5) have intermediate properties. Surprisingly, the differentiation-specific somatic histone H1.0 steadily increases from the normal astrocytes to the most transformed ones. As a whole, our results suggest that these three cell lines, together with the starting primary cells, constitute a potential model for studying glioma development.

Highlights

Brain cancers are complex and heterogeneous, and derive, in most cases, from glial cells [1,2,3,4,5]
In order to separate astrocytes from oligodendrocytes, we used a shaking procedure that takes advantage of the differences in adhesion of the various brain cell types, with astrocytes having a higher tendency to settle on the bottom of the culture flasks [37]
We found that primary astrocytes and A-GS1 doubled in about 60 h; while A-VV5 and A-FC6 showed doubling times of 36 and 25 h, respectively

Summary

Introduction

Brain cancers are complex and heterogeneous, and derive, in most cases, from glial cells [1,2,3,4,5]. These latter tumors are called gliomas, and are further subdivided into astrocytomas, oligodendrogliomas, ependymomas and glioastrocytomas [6,7]. On the basis of these findings, it is recommended to perform an “integrated diagnosis” of tumors, based on different criteria, such as histologic classification, the grade according to the World Health Organization (WHO), and molecular information [12]

Methods

Discussion

Conclusion