Survival and integration of transplanted postmitotic human neurons following experimental brain injury in immunocompetent rats.
OBJECTIVE
Limitations regarding cell homogeneity and survivability do not affect neuronlike hNT cells, which are derived from a human teratocarcinoma cell line (Ntera2) that differentiates into postmitotic neurons with exposure to retinoic acid. Because NT2N neurons survive longer than 1 year after transplantation into nude mice brains, the authors grafted these cells into the brains of immunocompetent rats following lateral fluid-percussion brain injury to determine the long-term survivability of NT2N cell grafts in cortices damaged by traumatic brain injury (TBI) and the therapeutic effect of NT2N neurons on cognitive and motor deficits.


METHODS
Seventy-two adult male Sprague-Dawley rats, each weighing between 340 and 370 g, were given an anesthetic agent and subjected to lateral fluid percussion brain injury of moderate severity (2.2-2.5 atm in 46 rats) or to surgery without TBI (shamoperation, 26 rats). Twenty-four hours postinjury, 10(5) NT2N cells (24 injured animals) or 3 microl of vehicle (22 injured and 14 control animals) was stereotactically implanted into the periinjured or control cerebral cortex. Motor function was assessed at weekly intervals and all animals were killed at 2 or 4 weeks after their posttraumatic learning ability was assessed using a Morris water maze paradigm. Viable NT2N grafts were routinely observed to extend human neural cell adhesion molecule-(MOC-1)immunoreactive processes into the periinjured cortex at 2 and 4 weeks posttransplantation, although no significant improvement in motor or cognitive function was noted. Inflammation identified around the transplant at both time points was assessed by immunohistochemical identification of macrophages (ED-1) and microglia (isolectin B4).


CONCLUSIONS
Long-term survival and integration of NT2N cells in the periinjured cortex of immunocompetent rats provides the researcher with an important cellular system that can be used to study maturation, regulation, and neurite outgrowth of transplanted neurons following TBI.
Department of Neurosurgery, University of Pennsylvania School of Medicine, Philadelphia 19104-6316, USA.
M F
MF
Philips
J K
JK
Muir
K E
KE
Saatman
R
R
Raghupathi
V M
VM
Lee
J Q
JQ
Trojanowski
T K
TK
McIntosh
D000818:Animals; D001930:Brain Injuries / Q000503:physiopathology / Q000601:surgery*; D002460:Cell Line; D002470:Cell Survival; D002540:Cerebral Cortex / Q000293:injuries / Q000503:physiopathology / Q000601:surgery; D003071:Cognition / Q000502:physiology; D004660:Encephalitis / Q000473:pathology; D005500:Follow-Up Studies; D006801:Humans; D007121:Immunocompetence; D007150:Immunohistochemistry; D007858:Learning / Q000502:physiology; D008264:Macrophages / Q000473:pathology; D008297:Male; D017628:Microglia / Q000473:pathology; D008938:Mitosis; D009068:Movement / Q000502:physiology; D019006:Neural Cell Adhesion Molecules / Q000032:analysis; D016501:Neurites / Q000502:physiology / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D014677:Pharmaceutical Vehicles; D051381:Rats; D017207:Rats, Sprague-Dawley; D013238:Stereotaxic Techniques; D014183:Transplantation, Heterologous
D019006:Neural Cell Adhesion Molecules; D014677:Pharmaceutical Vehicles