title: A neuroregenerative human ensheathing glia cell line with conditional rapid growth.
abstract: Ensheathing glia have been demonstrated to have neuroregenerative properties but this cell type from human sources has not been extensively studied because tissue samples are not easily obtained, primary cultures are slow growing, and human cell lines are not available. We previously isolated immortalized ensheathing glia by gene transfer of BMI1 and telomerase catalytic subunit into primary cultures derived from olfactory bulbs of an elderly human cadaver donor. These cells escape the replicative senescence characteristic of primary human cells while conserving antigenic and neuroregenerative properties of ensheathing glia, but their low proliferative rate in culture complicates their utility as cell models and their application for preclinical cell therapy experiments. In this study we describe the use of a conditional SV40 T antigen (TAg) transgene to generate human ensheathing glia cell lines, which are easy to maintain due to their robust growth in culture. Although these fast growing clones exhibited polyploid karyotypes frequently observed in cells immortalized by TAg, they did not acquire a transformed phenotype, all of them maintaining neuroregenerative capacity and antigenic markers typical of ensheathing glia. These markers were also retained even after elimination of the TAg transgene using Cre/LoxP technology, although the cells died shortly after, confirming that their survival depended on the presence of the immortalizing genes. We have also demonstrated here the feasibility of using these human cell lines in animal models by genetically marking the cells with GFP and implanting them into the injured spinal cord of immunosuppressed rats. Our conditionally immortalized human ensheathing glia cell lines will thus serve as useful tools for advancing cell therapy approaches and understanding neuroregenerative mechanisms of this unique cell type.
authors: Departamento de Biología Molecular, Universidad Autónoma de Madrid, Cantoblanco, Madrid, Spain.
authors: Vega
authors: V
authors: García-Escudero
authors: Ricardo
authors: R
authors: Gargini
authors: María Teresa
authors: MT
authors: Gallego-Hernández
authors: Ana
authors: A
authors: García-Gómez
authors: María Jesús
authors: MJ
authors: Martín-Bermejo
authors: Diana
authors: D
authors: Simón
authors: Alicia
authors: A
authors: Delicado
authors: María Teresa
authors: MT
authors: Moreno-Flores
authors: Jesús
authors: J
authors: Avila
authors: Filip
authors: F
authors: Lim
mesh_terms: D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D016923:Cell Death; D002460:Cell Line; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000473:pathology; D017690:Cell Transplantation; D006801:Humans; D007150:Immunohistochemistry; D007165:Immunosuppression Therapy; D019426:Integrases / Q000378:metabolism; D007621:Karyotyping; D051379:Mice; D008819:Mice, Nude; D009416:Nerve Regeneration / Q000502:physiology*; D009457:Neuroglia / Q000166:cytology* / Q000637:transplantation; D051381:Rats; D019076:Transgenes / Q000235:genetics; D014183:Transplantation, Heterologous
chemical_list: D000952:Antigens, Polyomavirus Transforming; C045073:Cre recombinase; D019426:Integrases