Neurons derived from a human teratocarcinoma cell line establish molecular and structural polarity following transplantation into the rodent brain.
Studies of neurons grafted into the brains of experimental animals have been limited by the lack of suitably homogeneous populations of neurons for transplantation. Here we describe the transplantation and survival of pure, postmitotic human neurons (NT2N cells) into the rat brain. NT2N cells were derived from a human teratocarcinoma line (NTera2/clone D1 or NT2 cells) in vitro by retinoic acid treatment. Approximately 5-10 x 10(4) NT2N cells (including previously frozen aliquots of NT2N cells) were injected into the neocortex, subjacent white matter, or hippocampus of adult (N = 51) or neonatal (N = 17) Sprague-Dawley rats. Cyclosporine (7-10 mg/kg) was administered daily to 13 adult rats for up to 12 weeks post-transplant prior to sacrifice. Untreated rats survived for up to 21 weeks post-transplant. Injection sites were serially sectioned and NT2N grafts were analyzed immunohistochemically using antibodies to diverse neuronal and glial proteins to assess the lineage of the grafted cells and their ability to establish molecular and structural polarity. NT2N cells transplanted into untreated adult and neonatal rat brains were committed exclusively to the neuronal phenotype and survived for as long as 8 weeks, although most were rejected after 4 weeks. However, cyclosporine prolonged survival of the NT2N grafts for up to 12 weeks. Further, grafted NT2N cells exhibited an asymmetric geometry (with long axons and simplified dendrites), as well as molecular polarity (with highly phosphorylated neurofilament proteins segregated in axons and microtubule associated protein 2 confined to perikarya and dendrites) by 4 weeks post-transplant. However, the grafted neurons did not become fully mature as evidenced by their failure to express the most highly phosphorylated heavy neurofilament proteins. Finally, previously frozen NT2N cells survived in the rat brain, and none of the grafts formed neoplasms. We conclude from these studies that transplanted NT2N cells represent a highly advantageous model system for studies of the developmental biology of neurons.
Trojanowski
J Q
JQ
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
Mantione
J R
JR
Lee
J H
JH
Seid
D P
DP
You
T
T
Inge
L J
LJ
Lee
V M
VM
D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000502:physiology*; D016764:Cell Polarity; D016922:Cellular Senescence; D016572:Cyclosporine / Q000494:pharmacology; D005260:Female; D006085:Graft Survival; D006801:Humans; D007150:Immunohistochemistry; D009368:Neoplasm Transplantation; D009474:Neurons / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013724:Teratoma / Q000378:metabolism* / Q000473:pathology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016572:Cyclosporine