鳥取臨床科学 2(1), 121-141, 2009

Immunocytochemical characterization of highly passaged primary astrocyte
cultures from autopsied aged human brain
Kaori Morimoto1, 2), Juri Horio1), Takeshi Kanesaka3), Kristina Lindholm4), Hiroyasu Akatsu3), Akira Hori3),
Kenji Okuda3), Takayuki Yamamoto3), Yong Shen4), Yoshihiro Konishi1)*1) Department of Clinical Research, National Tottori Medical Center, Tottori 689-0203, Japan
2) Department of Life Sciences, Faculty of Medicine, Tottori University, Yonago, Japan
3) Research Institute for Longevity Medicine, Fukushimura Hospital, Toyohashi, Japan
4) Haldeman Laboratory of Molecular and Cellular Neurobiology, Banner Sun Health Research Institute,
Sun City, AZ, USA*Correspondence: ykonishi@tottori-iryo.hosp.go.jp
AbstractAstrocytes have a wide variety of functions, unlike oligodendrocytes or ependymal cells that have a single
highly specialized function. Most of our understanding of astrocyte functions is derived from studies in vitro
utilizing astrocyte cultures established from neonatal and adult rodent brain and from fetal human brain.
However, to study the pathological roles of astrocytes in age-related neurological diseases in human, it iscrucial to establish in vitro systems derived from the adult human brain, predominantly from cases ofneurological disorders. In the past, methods have been developed to isolate viable glial cells from adult humanbrain tissue. We believe that human cells in the appropriate developmental, aging and disease state, and from
afflicted brain regions, would provide a more suitable in vitro model system for such studies. However, there
have been discrepancies in the literature as to the predominant type of cell in primary astrocyte cultures
established from adult human brain, particularly with multiple passages. Some reports say that the predominant
cell type is fibroblasts or fibroblast-like cells, and not cells of glial origin. Others contend that astrocytes
predominate. In the latter case, however, two different changes in GFAP expression in astrocytes weredemonstrated: stable and decreased expression with successive passages. Therefore, to examine whether
primary astrocytes are replaced with leptomeningeal fibroblasts and whether the GFAP immunoreactivity
changes during successive passages, we have conducted an immunocytochemical characterization of primary
astrocyte cultures established from aged human autopsied brain. As a result, more than 90% of the cultured
cells were thought to be astrocytes. Less than 10% were other types of cells, including fibroblasts, endothelial
cells, microglia, neurons and oligodendrocytes. Successive passages and even multiple cycles of freezing andthawing did not alter the antigenic properties of the cultured astrocytes. This indicates that primary cultures of
glial cells can be frozen for reposition in cell banks, distributed to laboratories around the world, and used as areliable source for glial cell research. Tottori J. Clin.Res. 2(1), 121-141, 2009

Key words: astrocytes, primary cultures, autopsy, glial acidic fibrillary protein (GFAP), cell bankbrain: neurons and glia. In the central nervous

Introduction
There are two major classes of cells in thesystem (CNS), glial cells are represented by three t121