Journal of Neuroscience, Vol 15, 5668-5680, Copyright © 1995 by Society for Neuroscience
CNS-derived neural progenitor cells for gene transfer of nerve growth factor to the adult rat brain: complete rescue of axotomized cholinergic neurons after transplantation into the septum
A Martinez-Serrano, C Lundberg, P Horellou, W Fischer, C Bentlage, K Campbell, RD McKay, J Mallet and A Bjorklund
Department of Medical Cell Research, University of Lund, Sweden.
A CNS-derived conditionally immortalized temperature-sensitive neural
progenitor (CINP) cell line was used to generate NGF-secreting cells
suitable for intracerebral transplantation. The cells were transduced by
repeated retroviral infection, using a vector containing the mouse NGF cDNA
under the control of the LTR promoter. Subcloning at the permissive
temperature (33 degrees C) identified a highly NGF-secreting clone
(NGF-CINP), which contained multiple copies of the transgene and released
NGF at a rate of 2 ng/hr/10(5) cells in vitro, both at 33 and 37 degrees C,
which was approximately 1 order of magnitude higher than what was possible
to achieve in the heterogeneously infected cell cultures. After
transplantation to the brain, the NGF-CINPs differentiated into cells with
a predominant glia-like morphology and migrated for a distance of 1-1.5 mm
from the implantation site into the surrounding host tissue, without any
signs of overgrowth and tumor formation. Grafts of NGF-CINP cells implanted
into the septum of adult rats with complete fimbria-fornix lesion blocked
over 90% of the cholinergic cell loss in the medial septum and grafts
placed in the intact striatum induced accumulation of low-affinity NGF
receptor positive fibers around the implantation site. Expression of the
NGF transgene in vivo was demonstrated by RT-PCR at 2 weeks after grafting.
It is concluded that the immortalized neural progenitors have a number of
advantageous properties that make them highly useful experimental tools for
gene transfer to the adult CNS.
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