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    • 9. 发明授权
    • Lineage-restricted neuronal precursors and methods of isolation
    • 谱系限制性神经元前体和分离方法
    • US06787353B1
    • 2004-09-07
    • US09109858
    • 1998-07-02
    • Mahendra S. RaoMargot Mayer-ProschelAnjali J. Kalyani
    • Mahendra S. RaoMargot Mayer-ProschelAnjali J. Kalyani
    • C12N508
    • C12N5/0623A61K35/12A61K48/00C12N2501/115C12N2501/13C12N2501/385
    • A self-renewing restricted stem cell population has been identified in developing (embryonic day 13.5) spinal cords that can differentiate into multiple neuronal phenotypes, but cannot differentiate into glial phenotypes. This neuronal-restricted precursor (NRP) expresses highly polysialated or embryonic neural cell adhesion molecule (E-NCAM) and is morphologically distinct from neuroepithelial stem cells (NEP cells) and spinal glial progenitors derived from embryonic day 10.5 spinal cord. NRP cells self renew over multiple passages in the presence of fibroblast growth factor (FGF) and neurotrophin 3 (NT-3) and express a characteristic subset of neuronal epitopes. When cultured in the presence of RA and the absence of FGF, NRP cells differentiate into GABAergic, glutaminergic, and cholinergic immunoreactive neurons. NRP cells can also be generated from multipotent NEP cells cultured from embryonic day 10.5 neural tubes. Clonal analysis shows that E-NCAM immunoreactive NRP cells arise from an NEP progenitor cell that generates other restricted CNS precursors. The NEP-derived E-NCAM immunoreactive cells undergo self renewal in defined medium and differentiate into multiple neuronal phenotypes in mass and clonal culture. Thus, a direct lineal relationship exists between multipotential NEP cells and more restricted neuronal precursor cells present in vivo at embryonic day 13.5 in the spinal cord. Methods for treating neurological diseases are also disclosed.
    • 在发育(胚胎期13.5)脊髓中已经鉴定出自我更新的限制性干细胞群,其可以分化成多个神经元表型,但不能分化成胶质表型。 这种神经元限制性前体(NRP)表达高度多聚化或胚胎神经细胞粘附分子(E-NCAM),并且在形态学上与神经上皮干细胞(NEP细胞)和源自胚胎第10.5天脊髓的脊髓神经祖细胞不同。 NRP细胞在成纤维细胞生长因子(FGF)和神经营养因子3(NT-3)的存在下自我更新多次,并表达神经元表位的特征亚群。 当在RA和无FGF的存在下培养时,NRP细胞分化为GABA能,谷氨酸能和胆碱能免疫反应性神经元。 NRP细胞还可以从从胚胎第10.5天神经管培养的多能NEP细胞产生。 克隆分析表明,E-NCAM免疫反应性NRP细胞来自产生其他受限制的CNS前体的NEP祖细胞。 NEP衍生的E-NCAM免疫反应性细胞在限定的培养基中进行自我更新,并在大量和克隆培养中分化成多个神经元表型。 因此,多潜能NEP细胞与脊髓中第13.5天体内存在的更多受限的神经元前体细胞之间存在直接的直线关系。 还公开了治疗神经疾病的方法。