title: Isolation and characterization of a spontaneously immortalized multipotent mesenchymal cell line derived from mouse subcutaneous adipose tissue.
abstract: The emerging field of tissue engineering and regenerative medicine is a multidisciplinary science that is based on the combination of a reliable source of stem cells, biomaterial scaffolds, and cytokine growth factors. Adult mesenchymal stem cells are considered important cells for applications in this field, and adipose tissue has revealed to be an excellent source of them. Indeed, adipose-derived stem cells (ASCs) can be easily isolated from the stromal vascular fraction (SVF) of adipose tissue. During the isolation and propagation of murine ASCs, we observed the appearance of a spontaneously immortalized cell clone, named m17.ASC. This clone has been propagated for more than 180 passages and stably expresses a variety of stemness markers, such as Sca-1, c-kit/CD117, CD44, CD106, islet-1, nestin, and nucleostemin. Furthermore, these cells can be induced to differentiate toward osteogenic, chondrogenic, adipogenic, and cardiogenic phenotypes. m17.ASC clone displays a normal karyotype and stable telomeres; it neither proliferates when plated in soft agar nor gives rise to tumors when injected subcutaneously in NOD/SCID-γ (null) mice. The analysis of gene expression highlighted transcriptional traits of SVF cells. m17.ASCs were genetically modified by lentiviral vectors carrying green fluorescent protein (GFP) as a marker transgene and efficiently engrafted in the liver, when injected in the spleen of NOD/SCID-γ (null) monocrotaline-treated mice. These results suggest that this non-tumorigenic spontaneously immortalized ASC line may represent a useful tool (cell model) for studying the differentiation mechanisms involved in tissue repair as well as a model for pharmacological/toxicological studies.
authors.affiliation: 1 Dipartimento di Scienze della Salute, Università del Piemonte Orientale , Novara, Italy .
authors.forename: Andrea
authors.initials: A
authors.lastname: Zamperone
authors.forename: Stefano
authors.initials: S
authors.lastname: Pietronave
authors.forename: Simone
authors.initials: S
authors.lastname: Merlin
authors.forename: Donato
authors.initials: D
authors.lastname: Colangelo
authors.forename: Gabriella
authors.initials: G
authors.lastname: Ranaldo
authors.forename: Enzo
authors.initials: E
authors.lastname: Medico
authors.forename: Federica
authors.initials: F
authors.lastname: Di Scipio
authors.forename: Giovanni Nicolao
authors.initials: GN
authors.lastname: Berta
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.forename: Maria
authors.initials: M
authors.lastname: Prat
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000818:Animals; D000950:Antigens, Ly / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D002999:Clone Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D053631:Interleukin Receptor Common gamma Subunit / Q000172:deficiency / Q000235:genetics; D059785:Karyotype; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D018613:Microscopy, Confocal; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D010011:Osteocytes / Q000166:cytology / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050151:Subcutaneous Fat / Q000166:cytology*
chemical_list: D000950:Antigens, Ly; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D049452:Green Fluorescent Proteins; D019009:Proto-Oncogene Proteins c-kit