Fu S-L, Ma Z-W, Yin L, Iannotti C, Lu P-H, Xu X-M
Department of Neurobiology, Shanghai Second Medical University, 280 South Chong-Qing Road, Shanghai 200025, PR China.
Neuroscience. 2005;135(3):851-62. doi: 10.1016/j.neuroscience.2005.05.049.
To determine whether neural precursor cells have region-specific growth properties, we compared the proliferation, mitogenicity, and differentiation of these cells isolated from the embryonic day 16 rat forebrain and spinal cord. Neural precursor cells isolated from both regions were cultured in growth medium supplemented with epidermal growth factor, basic fibroblast growth factor, or epidermal growth factor+basic fibroblast growth factor. Under all three conditions, both neural precursor cell populations proliferated for multiple passages. While spinal cord-derived neural precursor cells proliferated moderately faster in epidermal growth factor-enriched growth medium, brain-derived cells proliferated much faster in basic fibroblast growth factor-enriched growth medium. When exposed to both epidermal growth factor and basic fibroblast growth factor, the two neural precursor cell populations expanded and proliferated more rapidly than when exposed to a single factor, with brain-derived neural precursor cells expanding significantly faster than spinal cord-derived ones (P<0.0001). Differentiation studies showed that both neural precursor cell populations were multi-potent giving rise to neurons, astrocytes, and oligodendrocytes. However, neuronal differentiation from brain-derived neural precursor cells was greater than spinal cord-derived ones (11.95+/-5.00% vs 1.92+/-1.13%; passage 2). Further, the two neural precursor cell populations differentiated into a similar percentage of oligodendrocytes (brain: 8.66+/-5.85%; spinal cord: 7.69+/-3.91%; passage 2). Immunofluorescence and Western blot studies showed that neural precursor cells derived from both regions expressed receptors for basic fibroblast growth factor and epidermal growth factor. However, brain-derived neural precursor cells expressed higher levels of the two receptors than spinal cord-derived ones in growth medium containing epidermal growth factor+basic fibroblast growth factor. Thus, our results showed that neural precursor cells isolated from the two regions of the CNS have distinct properties and growth requirements. Identifying phenotypic differences between these neural precursor cell populations and their growth requirements should provide new insights into the development of cell therapies for region-specific neurological degenerative diseases.
为了确定神经前体细胞是否具有区域特异性生长特性,我们比较了从胚胎第16天大鼠前脑和脊髓分离的这些细胞的增殖、促有丝分裂活性和分化情况。从两个区域分离的神经前体细胞在补充有表皮生长因子、碱性成纤维细胞生长因子或表皮生长因子+碱性成纤维细胞生长因子的生长培养基中培养。在所有三种条件下,两个神经前体细胞群体均能传代多次进行增殖。虽然脊髓来源的神经前体细胞在富含表皮生长因子的生长培养基中增殖速度稍快,但脑来源的细胞在富含碱性成纤维细胞生长因子的生长培养基中增殖速度更快。当同时暴露于表皮生长因子和碱性成纤维细胞生长因子时,两个神经前体细胞群体比暴露于单一因子时扩增和增殖得更快,其中脑来源的神经前体细胞扩增速度明显快于脊髓来源的神经前体细胞(P<0.0001)。分化研究表明,两个神经前体细胞群体都是多能的,可产生神经元、星形胶质细胞和少突胶质细胞。然而,脑来源的神经前体细胞向神经元的分化程度高于脊髓来源的神经前体细胞(第2代时为11.95±5.00%对1.92±1.13%)。此外,两个神经前体细胞群体分化为少突胶质细胞的比例相似(脑:8.66±5.85%;脊髓:7.69±3.91%;第2代)。免疫荧光和蛋白质印迹研究表明,来自两个区域的神经前体细胞均表达碱性成纤维细胞生长因子和表皮生长因子的受体。然而,在含有表皮生长因子+碱性成纤维细胞生长因子的生长培养基中,脑来源的神经前体细胞表达这两种受体的水平高于脊髓来源的神经前体细胞。因此,我们的结果表明,从CNS的两个区域分离的神经前体细胞具有不同的特性和生长需求。确定这些神经前体细胞群体之间的表型差异及其生长需求,应为针对区域特异性神经退行性疾病的细胞治疗开发提供新的见解。