Horiuchi Makoto, Tomooka Yasuhiro
Department of Biological Science and Technology, Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda City, Chiba 278-8510, Japan.
Neurosci Res. 2005 Oct;53(2):104-15. doi: 10.1016/j.neures.2005.06.007.
In the present study, a clonal astrocyte progenitor cell line derived from p53-deficient fetal brains, named FBD-104, was characterized in monolayer and suspension culture. In monolayer culture with medium containing 10% serum, FBD-104 cells expressed some markers of astrocytes, such as glial fibrillary acidic protein (GFAP), S100beta, and glutamate aspartate transporter (GLAST). They never expressed any markers of neurons or oligodendrocytes. Thus the cell line appears to be restricted to the astroglial lineage. However, in suspension culture in serum-free medium supplemented with EGF and FGF2, FBD-104 cells proliferated and formed neurospheres expressing mRNAs for Mash1 and Math3, generating cells expressing neuron specific beta-III tubulin. Re-plating the spheres onto an adhesive substrate and withdrawal of the growth factors induced the expression of mRNAs for NeuroD and Olig2 and generated more beta-III tubulin-positive cells. The present study demonstrated that neurosphere culture is an efficient method to induce neurogenesis from the astrocyte progenitor cell line FBD-104. We also determined that pretreatment with FGF2 caused a significant increase in yield of neurospheres. Thus, the FBD-104 line is an interesting in vitro model to study effect of trophic factors and adhesive substrates on lineage determination of neural progenitor cells.
在本研究中，对一种源自p53基因缺陷型胎脑的克隆性星形胶质细胞祖细胞系（命名为FBD - 104）进行了单层培养和悬浮培养特性分析。在含有10%血清的培养基中进行单层培养时，FBD - 104细胞表达了一些星形胶质细胞的标志物，如胶质纤维酸性蛋白（GFAP）、S100β和谷氨酸天冬氨酸转运体（GLAST）。它们从未表达任何神经元或少突胶质细胞的标志物。因此，该细胞系似乎局限于星形胶质细胞谱系。然而，在补充有表皮生长因子（EGF）和碱性成纤维细胞生长因子2（FGF2）的无血清培养基中进行悬浮培养时，FBD - 104细胞增殖并形成表达Mash1和Math3 mRNA的神经球，产生表达神经元特异性β-III微管蛋白的细胞。将这些神经球重新接种到黏附性底物上并撤除生长因子，可诱导NeuroD和Olig2 mRNA的表达，并产生更多β-III微管蛋白阳性细胞。本研究表明，神经球培养是一种从星形胶质细胞祖细胞系FBD - 104诱导神经发生的有效方法。我们还确定，用FGF2预处理可显著提高神经球的产量。因此，FBD - 104细胞系是研究营养因子和黏附性底物对神经祖细胞谱系决定作用的一个有趣的体外模型。
