Hall Peter E, Lathia Justin D, Caldwell Maeve A, Ffrench-Constant Charles
Department of Pathology, University of Cambridge, Cambridge, UK.
BMC Neurosci. 2008 Jul 23;9:71. doi: 10.1186/1471-2202-9-71.
Human neural stem cells (hNSC) have the potential to provide novel cell-based therapies for neurodegenerative conditions such as multiple sclerosis and Parkinson's disease. In order to realise this goal, protocols need to be developed that allow for large quantities of hNSC to be cultured efficiently. As such, it is important to identify factors which enhance the growth of hNSC. In vivo, stem cells reside in distinct microenvironments or niches that are responsible for the maintenance of stem cell populations. A common feature of niches is the presence of the extracellular matrix molecule, laminin. Therefore, this study investigated the effect of exogenous laminin on hNSC growth.
To measure hNSC growth, we established culture conditions using B27-supplemented medium that enable neurospheres to grow from human neural cells plated at clonal densities. Limiting dilution assays confirmed that neurospheres were derived from single cells at these densities. Laminin was found to increase hNSC numbers as measured by this neurosphere formation. The effect of laminin was to augment the proliferation/survival of the hNSC, rather than promoting the undifferentiated state. In agreement, apoptosis was reduced in dissociated neurospheres by laminin in an integrin beta1-dependent manner.
The addition of laminin to the culture medium enhances the growth of hNSC, and may therefore aid their large-scale production.
人类神经干细胞（hNSC）有潜力为诸如多发性硬化症和帕金森病等神经退行性疾病提供新型的基于细胞的治疗方法。为实现这一目标，需要开发能够高效培养大量hNSC的方案。因此，识别促进hNSC生长的因素很重要。在体内，干细胞存在于负责维持干细胞群体的独特微环境或生态位中。生态位的一个共同特征是细胞外基质分子层粘连蛋白的存在。因此，本研究调查了外源性层粘连蛋白对hNSC生长的影响。
为了测量hNSC的生长，我们使用添加了B27的培养基建立了培养条件，该条件能使神经球从以克隆密度接种的人类神经细胞生长出来。有限稀释分析证实，在这些密度下神经球来源于单细胞。通过这种神经球形成测定发现，层粘连蛋白可增加hNSC数量。层粘连蛋白的作用是增强hNSC的增殖/存活，而不是促进其未分化状态。与此一致的是，层粘连蛋白以整合素β1依赖的方式减少了解离的神经球中的细胞凋亡。
向培养基中添加层粘连蛋白可增强hNSC的生长，因此可能有助于其大规模生产。
