Neural stem cell transplantation in cold lesion: a novel approach for the investigation of brain trauma and repair.
We developed a new neural transplantation protocol for the investigation of the repair of brain trauma. Cortical lesion was induced by touching a cold (-60 degrees C) metal stamp to the dura over the forelimb motor cortex of adult rats. The procedure caused a localized lesion and the animals developed a significant motor deficit, which was monitored throughout the protocol. Six days later the animals received embryonic neural stem cells in the penumbra of the lesion. The donor cells were freshly isolated from E14 rat embryos, had a high viability, and expressed the stem cell marker nestin. A further 6 days later the survival and differentiation of the grafted cells were investigated by immunohistochemistry. The majority of the surviving grafted cells were found in the lesion and they did not express lineage-specific markers. Only 10% of all surviving transplanted cells were located in the penumbra. These cells had an astrocytic phenotype and expressed glial fibrillary acidic protein. A few cells expressed neural or oligodendrocytic markers. In conclusion, we established a novel neural transplantation protocol, which focuses on cortical brain trauma. The model is a combination of surgical, neurological and histological approaches, all adapted to each other to make a reliable and reproducible experimental model.
Lacza
Zsombor
Z
Department of Physiology/Pharmacology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USA. zlacza@mac.com
Horváth
Eszter
E
Busija
David W
DW
D000165:Acridine Orange / Q000493:pharmacokinetics; D000818:Animals; D001930:Brain Injuries / Q000628:therapy*; D016380:Brain Tissue Transplantation; D001973:Bromodeoxyuridine / Q000493:pharmacokinetics; D002454:Cell Differentiation / Q000502:physiology; D003080:Cold Temperature / Q000009:adverse effects*; D004622:Embryo, Mammalian; D011464:Epoprostenol / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D004996:Ethidium / Q000493:pharmacokinetics; D005260:Female; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D007202:Indicators and Reagents / Q000493:pharmacokinetics; D007381:Intermediate Filament Proteins / Q000378:metabolism; D008297:Male; D009044:Motor Cortex / Q000378:metabolism / Q000637:transplantation; D009419:Nerve Tissue Proteins; D064231:Nestin; D009930:Organic Chemicals; D011247:Pregnancy; D011838:Radiation-Sensitizing Agents / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D033581:Stem Cell Transplantation / Q000379:methods*; D013997:Time Factors; D014945:Wound Healing
D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D007202:Indicators and Reagents; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D009930:Organic Chemicals; D011838:Radiation-Sensitizing Agents; C405903:TO-PRO-1; C068518:TTC 909; D011464:Epoprostenol; D004996:Ethidium; D000165:Acridine Orange; D001973:Bromodeoxyuridine