DIFFERENTIATION OF BIPOTENTIAL GLIAL PRECURSORS INTO OLIGODENDROCYTES IS PROMOTED BY INTERACTION WITH TYPE-1 ASTROCYTES IN CEREBELLAR CULTURES
DIFFERENTIATION OF BIPOTENTIAL GLIAL PRECURSORS INTO OLIGODENDROCYTES IS PROMOTED BY INTERACTION WITH TYPE-1 ASTROCYTES IN CEREBELLAR CULTURES
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DOI：
10.1073/pnas.85.16.6167
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发表时间：
1988-08-01
影响因子：
11.1
通讯作者：
LEVI, G
中科院分区：
文献类型：
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作者：
ALOISI, F;AGRESTI, C;LEVI, G
用LB 1抗体标记原代培养的大鼠小脑，观察了少突胶质细胞（OL）和2型星形胶质细胞（AS）的分化（其结合存在于神经胶质前体、2型AS和未成熟OL中的表面二唾液酸神经节苷脂GD 3）、O 4（未成熟和成熟OL与表面硫脂结合的标志物）、抗半乳糖苷（GalCer，OL的标志物）和抗神经胶质细胞酸性蛋白（GFAP，AS的标志物）。接种后2小时，几乎检测不到任何LB 1+、GFAP+细胞，40%的0 4+细胞为GalCer+，且0 4+细胞均为GFAP+。在培养细胞时，以1 × 10 - 4的密度铺板。105个细胞/cm 2的胎牛血清中，大多数LB 1+前体分化为2型AS，即使他们中的大多数已经表达O 4抗原。因此，在培养中，大多数2型AS似乎来自体内分化为OL的祖细胞。在更高密度的培养物中（2.5 × 10 - 4），105个细胞/cm 2），然而，许多前体细胞分化成GalCer+ OL，而不是分化成AS。作为负责高密度培养物中神经胶质前体行为的信号的可能来源，我们将注意力集中在培养物中最丰富的细胞类型1型AS上。我们发现，在低密度培养物中保持5-7天的条件培养基中的1型AS，前体细胞的增殖得到加强，并阻止其分化为OL或AS。相反，当小脑细胞与纯化培养物中分离的1型AS共培养时，不仅前体细胞比对照培养物增殖更多，而且它们中的更大比例分化为GalCer+ OL。总之，1型AS似乎通过直接的细胞-细胞相互作用促进双能胶质前体分化为OL。1型AS对LB 1+和O 4+前体分化的影响也得到了神经胶质皮层培养实验的支持。
The differentiation of bipotential precursors of oligodendrocytes (OL) and type-2 astrocytes (AS) was followed in primary cultures from 8-day postnatal rat cerebellum by labeling the cells with the antibodies LB1 (which binds to the surface disialoganglioside GD3 present in glial precursors, type-2 AS, and immature OL), O4 (a marker of immature and mature OL binding to surface sulfatide), anti-galactocerebroside (GalCer, a marker of OL), and anti-glial fibrillary acidic protein (GFAP, a marker of AS). Two hours after plating, hardly any LB1+, GFAP+ cells were detectable, 40% of the O4+ cells were GalCer+, and none of the O4+ cells were GFAP+. Upon culturing cells plated at a density of 1 .times. 105 cells per cm2 in the presence of fetal calf serum, most of the LB1+ precursors differentiated into type-2 AS, even if most of them had already expressed the O4 antigen. Thus, in culture, most type-2 AS seem to derive from progenitor cells that were differentiating in vivo into OL. In higher density cultures (2.5 .times. 105 cells per cm2), however, many precursors differentiated into GalCer+ OL, rather than into AS. As a possible source of the signals responsible for the behavior of the glial precursors in high-density cultures, we focused our attention on type 1-AS, the most abundant cell type in the cultures. We found that, in low-density cultures maintained for 5-7 days in a medium conditioned by type-1 AS, the proliferation of the precursors was enhanced and their differentiation into OL or AS was prevented. In contrast, when cerebellar cells were coplated with type-1 AS dissociated from purified cultures, not only did the precursors proliferate more than in control cultures, but also a larger proportion of them differentiated into GalCer+ OL. In conclusion, type-1 AS appear to facilitate the differentiation of bipotential glial precursors into OL through direct cell-cell interactions. The influence of type-1 AS on the differentiation of the LB1+ and O4+ precursors is supported also by experiments with glial cortical cultures.