Shihabuddin L S, Ray J, Gage F H
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
Exp Neurol. 1997 Dec;148(2):577-86. doi: 10.1006/exnr.1997.6697.
The adult rat brain contains progenitor cells that can be induced to proliferate in vitro in response to FGF-2. In the present study we explored whether similar progenitor cells can be cultured from different levels (cervical, thoracic, lumbar, and sacral) of adult rat spinal cord and whether they give rise to neurons and glia as well as spinal cord-specific neurons (e.g., motoneurons). Cervical, thoracic, lumbar, and sacral areas of adult rat spinal cord (>3 months old) were microdissected and neural progenitors were isolated and cultured in serum-free medium containing FGF-2 (20 ng/ml) through multiple passages. Although all areas generated rapidly proliferating cells, the cultures were heterogeneous in nature and cell morphology varied within a given area as well as between areas. A percentage of cells from all areas of the spinal cord differentiate into cells displaying antigenic properties of neuronal, astroglial, and oligodendroglial lineages; however, the majority of cells from all regions expressed the immature proliferating progenitor marker vimentin. In established multipassage cultures, a few large, neuron-like cells expressed immunoreactivity for p75NGFr and did not express GFAP. These cells may be motoneurons. These results demonstrate that FGF-2 is mitogenic for progenitor cells from adult rat spinal cord that have the potential to give rise to glia and neurons including motoneurons.
成年大鼠大脑中含有祖细胞,这些祖细胞可在体外被诱导响应FGF-2而增殖。在本研究中,我们探究了是否可以从成年大鼠脊髓的不同节段(颈段、胸段、腰段和骶段)培养出类似的祖细胞,以及它们是否能分化为神经元、神经胶质细胞以及脊髓特异性神经元(如运动神经元)。对成年大鼠(大于3个月龄)脊髓的颈段、胸段、腰段和骶段进行显微解剖,分离神经祖细胞,并在含有FGF-2(20 ng/ml)的无血清培养基中进行多次传代培养。尽管所有节段都能产生快速增殖的细胞,但培养物本质上是异质性的,给定节段内以及不同节段之间的细胞形态各不相同。脊髓所有节段的一定比例的细胞分化为具有神经元、星形胶质细胞和少突胶质细胞系抗原特性的细胞;然而,所有区域的大多数细胞都表达未成熟的增殖祖细胞标志物波形蛋白。在已建立的多次传代培养物中,一些大的、神经元样细胞对p75NGFr呈免疫反应性,且不表达GFAP。这些细胞可能是运动神经元。这些结果表明,FGF-2对成年大鼠脊髓中的祖细胞具有促有丝分裂作用,这些祖细胞有可能分化为神经胶质细胞和神经元,包括运动神经元。