Kukekov V G, Laywell E D, Suslov O, Davies K, Scheffler B, Thomas L B, O'Brien T F, Kusakabe M, Steindler D A
Department of Anatomy and Neurobiology, University of Tennessee at Memphis, Memphis, Tennessee, 38163, USA.
Exp Neurol. 1999 Apr;156(2):333-44. doi: 10.1006/exnr.1999.7028.
Recent in vitro studies have shown that the periventricular subependymal zone (SEZ) of the rodent brain is capable of de novo generation of neurons and glia. There is less information available on neurogenesis in the adult human brain, and no study has shown the clonal generation of neurons and glia from in vitro-generated "neurospheres." Here we describe the isolation of proliferative stem/progenitor cells within neurospheres from two different regions, the SEZ and the hippocampus, from surgical biopsy specimens of adult (24-57 years) human brain. Using light and electron microscopy; immunocytochemistry for a variety of neuronal, glial, and developmental (including extracellular matrix; ECM) markers; and the reverse transcriptase polymerase chain reaction to demonstrate different gene transcripts found in neurospheres, it is shown that the adult human brain harbors a complex population of stem/progenitor cells that can generate neuronal and glial progeny under particular in vitro growth conditions. These methods also show that these neurospheres contain both neurons and glia and demonstrate regional similarities at the mRNA level, indicating common stem/progenitor cell types within two different neurogenic regions of the adult human brain. In addition to the synthesis of developmentally regulated molecules such as the ECM protein tenascin-C, a variety of other genes (e.g., Pax 6) and proteins (e.g. , Bcl-2) involved in cell survival and differentiation are expressed by adult human brain neurospheres.
最近的体外研究表明，啮齿动物脑室内周围的室管膜下区（SEZ）能够从头生成神经元和神经胶质细胞。关于成人人脑中神经发生的信息较少，且尚无研究表明体外生成的“神经球”能克隆生成神经元和神经胶质细胞。在此，我们描述了从成人（24 - 57岁）人脑手术活检标本的两个不同区域，即SEZ和海马体中分离神经球内增殖性干细胞/祖细胞的方法。通过光学和电子显微镜；针对多种神经元、神经胶质和发育（包括细胞外基质；ECM）标志物的免疫细胞化学；以及逆转录聚合酶链反应来证明神经球中发现的不同基因转录本，结果表明，成人人脑中有一群复杂的干细胞/祖细胞，在特定的体外生长条件下能够生成神经元和神经胶质后代。这些方法还表明，这些神经球同时包含神经元和神经胶质细胞，并在mRNA水平显示出区域相似性，这表明成人人脑两个不同神经发生区域内存在共同的干细胞/祖细胞类型。除了合成发育调控分子，如ECM蛋白腱生蛋白-C外，成人人脑神经球还表达多种参与细胞存活和分化的其他基因（如Pax 6）和蛋白质（如Bcl-2）。
