Widera Darius, Grimm Wolf-Dieter, Moebius Jeannette M, Mikenberg Ilja, Piechaczek Christoph, Gassmann Georg, Wolff Natascha A, Thévenod Frank, Kaltschmidt Christian, Kaltschmidt Barbara
Institute of Neurobiochemistry, University of Witten, Herdecke, Witten, Germany.
Stem Cells Dev. 2007 Jun;16(3):447-60. doi: 10.1089/scd.2006.0068.
Neural stem cells (NSCs) are potential sources for cell therapy of neurodegenerative diseases and for drug screening. Despite their potential benefits, ethical and practical considerations limit the application of NSCs derived from human embryonic stem cells (ES) or adult brain tissue. Thus, alternative sources are required to satisfy the criteria of ready accessibility, rapid expansion in chemically defined media and reliable induction to a neuronal fate. We isolated somatic stem cells from the human periodontium that were collected during minimally invasive periodontal access flap surgery as part of guided tissue regeneration therapy. These cells could be propagated as neurospheres in serum-free medium, which underscores their cranial neural crest cell origin. Culture in the presence of epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2) under serum-free conditions resulted in large numbers of nestin-positive/Sox-2-positive NSCs. These periodontium-derived (pd) NSCs are highly proliferative and migrate in response to chemokines that have been described as inducing NSC migration. We used immunocytochemical techniques and RT-PCR analysis to assess neural differentiation after treatment of the expanded cells with a novel induction medium. Adherence to substrate, growth factor deprivation, and retinoic acid treatment led to the acquisition of neuronal morphology and stable expression of markers of neuronal differentiation by more than 90% of the cells. Thus, our novel method might provide nearly limitless numbers of neuronal precursors from a readily accessible autologous adult human source, which could be used as a platform for further experimental studies and has potential therapeutic implications.
神经干细胞（NSCs）是神经退行性疾病细胞治疗和药物筛选的潜在来源。尽管它们有潜在益处，但伦理和实际考虑限制了源自人类胚胎干细胞（ES）或成体脑组织的神经干细胞的应用。因此，需要替代来源来满足易于获取、在化学成分明确的培养基中快速扩增以及可靠诱导分化为神经元命运的标准。我们从人类牙周组织中分离出体细胞干细胞，这些细胞是在微创牙周翻瓣手术中作为引导组织再生治疗的一部分收集的。这些细胞可以在无血清培养基中作为神经球进行增殖，这突出了它们源自颅神经嵴细胞。在无血清条件下，在表皮生长因子（EGF）和成纤维细胞生长因子-2（FGF-2）存在的情况下进行培养，可产生大量巢蛋白阳性/ Sox-2阳性神经干细胞。这些源自牙周组织（pd）的神经干细胞具有高度增殖能力，并能响应已被描述为诱导神经干细胞迁移的趋化因子而迁移。我们使用免疫细胞化学技术和逆转录-聚合酶链反应（RT-PCR）分析来评估用新型诱导培养基处理扩增细胞后的神经分化情况。细胞贴壁、生长因子剥夺和视黄酸处理导致超过90%的细胞获得神经元形态并稳定表达神经元分化标志物。因此，我们的新方法可能从易于获取的自体成人来源提供几乎无限数量的神经元前体细胞，这可作为进一步实验研究平台，并具有潜在治疗意义。
