Serum and transforming growth factor beta regulate glial fibrillary acidic protein in serum-free-derived mouse embryo cells.

Abstract

Serum-free mouse embryo (SFME) cells, derived in medium in which serum is replaced with growth factors and other supplements, display distinctive properties: (i) SFME cells do not lose proliferative potential or show gross chromosomal aberration upon extended culture, (ii) these cells depend on epidermal growth factor for survival; and (ii) SFME cell proliferation is reversibly inhibited by serum. Treatment of SFME cells with serum or transforming growth factor j3 led to the appearance of glial fibrillary acidic protein, a specific marker for astrocytes. The appearance of glial fibrillary acidic protein in cultures was reversed upon removal of transforming growth factor (3 or serum. Cells with properties similar to SFME cells were also isolated from adult mouse brain. These results suggest a role for transforming growth factor (3 in astrocyte differentiation in developing organisms and in response to injury and identify the cell type that has the unusual properties of SFME cells.Serum-free mouse embryo (SFME) cells initiated and maintained multipassage in a rich basal nutrient medium supplemented with insulin, transferrin, epidermal growth factor (EGF), high-density lipoprotein, and fibronectin display several distinctive properties (1)(2)(3)(4)(5)(6). Mouse embryo cells derived in conventional serum-supplemented medium undergo a period in which proliferative potential is lost or reduced, and then genetically altered cells with indefinite proliferative potential emerge (2,7,8). SFME cells do not exhibit any loss of proliferative potential or gross chromosomal aberration and maintain a relatively stable karyotype when cultured for >10 times the population doublings achieved with karyotypically normal mouse embryo cells in serum-containing medium. Although the period in which reduced growth rate can be detected may be minimal or absent in cultures in serumcontaining medium maintained in a passaging protocol involving high plating densities, the protocols used to derive SFME cells lead to a well-defined period of limited proliferative potential in cultures maintained in serum-containing medium (2, 7) that is not seen when the cells are cultured in serum-free medium. SFME cells also depend on EGF for survival, and cycloheximide or actinomycin D prevents death from EGF deprivation (5), suggesting that cell death requires the synthesis of RNA and protein, a phenomenon of programmed cell death similar to that reported for neuronal cell death in the absence of nerve growth factor (NGF) (9). In addition, proliferation of SFME cells is reversibly inhibited by serum or platelet-free plasma (1, 4, 6), and thus these cells would not be propagated under conventional culture conditions using serum-supplemented medium. As for most mouse embryo cell lines, the precise nature of the cells giving rise to the continuously growing SFME cultures was unknown. The unusual characteristics of SFME cells led us to attempt to determine the cell type represented by these cells.We found that treatment of SFME cells with serum or We a...

RETRACTED ARTICLE: Mesenchymal stem cells and neural crest stem cells from adult bone marrow: characterization of their surprising similarities and differences

“…Noteworthy, the regulation of nestin expression was different between MSC and NCSC as serum was able to repress this expression only in MSC, while no serum effect was observed on NCSC. A similar phenomenon has already been described for glial precursor cells in which serum (through TGFβ factors) inhibited nestin expression [15]. Moreover, we previously demonstrated that thrombin was able to upregulate nestin expression by radial glial cells, but decreased the nestin expression in MSC cell population from bone marrow, indicating that the regulation of nestin expression could be finely modulated in various conditions and/or cell types [12].…”

Characterization of Progenitor-Cell-Specific Genes Identified by Subtractive Suppression Hybridization

“…It has been reported that a mouse stem cell line (SFME) when stimulated by transforming growth factor ß (TGF-ß) or serum begins to express cystatin C, as well as a mouse homologue of the H2-4 clone detected in this paper and an astrocyte marker, GFAP [Sakai et al, 1990;Solem et al, 1990;Weisz et al, 1993]. Our results showing upregulation of both cystatin C and H2-4 in GRIP cells compared to NEP cells are consistent with these reports.…”

Differentiation of Serum-Free Mouse Embryo Cells into an Astrocytic Lineage is Associated with the Asymmetric Production of Early Neural, Neuronal and Glial Markers

“…It was also found that differentiation of these cells into an astrocytic lineage is associated with asymmetric production of these markers, rather than by acquiring an astrocytic marker upon differentiation. SFME cells were originally derived from a 16-d-old whole mouse embryo 29) and have been documented to increase GFAP, the astrocyte intermediate filament marker protein, following the addition of serum, TGF-b, 56,57) LIF 58) or BMPs 59) or following the addition of LIF plus BMP2. 60) These cells offer advantages for studying cell differentiation because they serve as progenitor cells and proliferate without senescence in vitro, 34,59) and have long been considered to differentiate into only an astrocytic lineage.…”