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 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.
Martínez-Serrano
A
A
Department of Medical Cell Research, University of Lund, Sweden.
Lundberg
C
C
Horellou
P
P
Fischer
W
W
Bentlage
C
C
Campbell
K
K
McKay
R D
RD
Mallet
J
J
Björklund
A
A
D000818:Animals; D001369:Axons / Q000502:physiology; D001483:Base Sequence; D001921:Brain / Q000502:physiology*; D002460:Cell Line; D002490:Central Nervous System / Q000166:cytology*; D005260:Female; D018014:Gene Transfer Techniques; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D009474:Neurons / Q000502:physiology* / Q000637:transplantation; D010275:Parasympathetic Nervous System / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012688:Septum Pellucidum / Q000502:physiology; D013234:Stem Cells / Q000502:physiology*
D015335:Molecular Probes; D009414:Nerve Growth Factors