A new method for the rapid and long term growth of human neural precursor cells
A new method for the rapid and long term growth of human neural precursor cells
复制标题
DOI：
10.1016/s0165-0270(98)00126-5
复制
发表时间：
1998-12-01
影响因子：
3
通讯作者：
Caldwell, MA
中科院分区：
文献类型：
--
作者：
Svendsen, CN;ter Borg, MG;Caldwell, MA
人类神经组织的可靠来源对神经科学家和临床神经移植试验都具有巨大的实用价值。在这项研究中，从发育中的人类皮质中分离出人的前体细胞，并在表皮和成纤维细胞生长因子-2存在的情况下，在培养中以球形簇的形式生长。使用传统的传代技术和培养液，生长速度极其缓慢，细胞总数只能扩大12倍。然而，当完整的球体被分割成几个季度，而不是机械地分离时，细胞与细胞之间的接触保持不变，细胞损伤最小化，这使得每个单独的季度能够快速和持续地增长。使用这种方法，在不到200天的时间里，我们的前体细胞数量增加了150万倍。通过暴露在底物中进行分化后，细胞从球体中迁移出来，形成星形胶质细胞和神经元的单层。当整个球体分化时，这些人类神经前体细胞在传代后期没有发育出少突胶质细胞。这种简单而新颖的培养方法允许大量未转化的人类神经前体细胞快速扩增，可能在药物发现、体外基因治疗和临床神经移植中使用。(C)1998 Elsevier Science B.V.保留所有权利。
A reliable source of human neural tissue would be of immense practical value to both neuroscientists and clinical neural transplantation trials. In this study, human precursor cells were isolated from the developing human cortex and, in the presence of both epidermal and fibroblast growth factor-2, grew in culture as sphere shaped clusters. Using traditional passaging techniques and culture mediums the rate of growth was extremely slow, and only a 12-fold expansion in total cell number could be achieved. However, when intact spheres were sectioned into quarters, rather than mechanically dissociated, cell-cell contacts were maintained and cellular trauma minimised which permitted the rapid and continual growth of each individual quarter. Using this method we have achieved a 1.5 million-fold increase in precursor cell number over a period of less than 200 days. Upon differentiation by exposure to a substrate, cells migrated out from the spheres and formed a monolayer of astrocytes and neurons. No oligodendrocytes were found to develop from these human neural precursor cells at late passages when whole spheres were differentiated. This simple and novel culture method allows the rapid expansion of large numbers of non-transformed human neural precursor cells which may be of use in drug discovery, ex vivo gene therapy and clinical neural transplantation. (C) 1998 Elsevier Science B.V. All rights reserved.