Siebzehnrubl Florian A, Steindler Dennis A
Department of Neurosurgery, McKnight Brain Institute, University of Florida, Gainesville, FL, USA.
Methods Mol Biol. 2013;1059:79-86. doi: 10.1007/978-1-62703-574-3_7.
Adult neural precursor cells are an essential part of the brain, and a focus of two decades of intense research (Ming and Song, Neuron 70:687-702, 2011). Even though adult human stem/progenitor cells have been identified early on (Kirschenbaum et al., Cereb Cortex 4:576-589, 1994; Eriksson et al., Nat Med 4:1313-1317, 1998), progress in the field of adult human neurogenesis has been slow. The reasons for this may be more advanced neighboring fields of pluripotent stem cell research, and lacking study material as well as well-established and standardized protocols. Furthermore, adult precursor cells in humans seem to have greater potential than in rodents (Walton et al., Development 133:3671-3681, 2006). This may be attributed to species differences in astrocyte development and diversity (Oberheim et al., Neurosci 29:3276-3287, 2009). In this chapter, we provide a guideline for adult human brain tissue dissociation, be it from biopsy or autopsy specimens. This is by no means the only way of culturing adult neural precursor cells, but it may help in streamlining research on this fascinating topic, as well as help introducing others into this field. We describe our methodology for establishing and maintaining long-term cultures from white and grey matter, as well as a simple protocol for differentiating these cells.
成体神经前体细胞是大脑的重要组成部分，也是二十年来深入研究的焦点（Ming和Song，《神经元》70:687 - 702，2011年）。尽管成人的干细胞/祖细胞早在早期就已被鉴定出来（Kirschenbaum等人，《大脑皮层》4:576 - 589，1994年；Eriksson等人，《自然医学》4:1313 - 1317，1998年），但成人神经发生领域的进展一直缓慢。其原因可能是多能干细胞研究的相邻领域更为先进，且缺乏研究材料以及完善和标准化的方案。此外，人类的成体前体细胞似乎比啮齿动物具有更大的潜力（Walton等人，《发育》133:3671 - 3681，2006年）。这可能归因于星形胶质细胞发育和多样性的物种差异（Oberheim等人，《神经科学》29:3276 - 3287，2009年）。在本章中，我们提供了一份关于成人脑组织解离的指南，无论是来自活检还是尸检标本。这绝不是培养成体神经前体细胞的唯一方法，但它可能有助于简化对这一迷人主题的研究，并帮助其他人进入该领域。我们描述了从白质和灰质建立和维持长期培养物的方法，以及这些细胞分化的简单方案。
