title: Fluorescent Immortalized Human Adipose Derived Stromal Cells (hASCs-TS/GFP+) for Studying Cell Drug Delivery Mediated by Microvesicles.
abstract: BACKGROUND
  A new tool for the drug delivery is based on the use of Mesenchymal Stromal Cells (MSCs) loaded in vitro with anti-cancer drugs. Unfortunately, the restricted lifespan of MSCs represents a significant limitation to produce them in high amounts and for long time studies. Immortalized MSCs from adipose tissue (hASCs) have been generated as good source of cells with stable features. These cells could improve the development of standardized procedures for both in vitro and preclinical studies. Furthermore they facilitate procedures for preparing large amounts of secretome containing microvesicles (MVs).
  
  
  METHODS
  We used human adipose tissue derived MSCs immortalized with hTERT+SV40 (TS) genes and transfected with GFP (hASCs-TS/GFP+). This line was investigated for its ability to uptake and release anticancer drugs. Microvesicles associated to paclitaxel (MVs/PTX) were isolated, quantified, and tested on pancreatic cancer cells.
  
  
  RESULTS
  The line hASCs-TS/GFP+ maintained the main mesenchymal characters and was able to uptake and release, in active form, both paclitaxel and gemcitabine. From paclitaxel loaded hASCs-TS/GFP+ cells were isolated microvesicles in sufficient amount to inhibit "in vitro" the proliferation of pancreatic tumor cells.
  
  
  CONCLUSIONS
  Our study suggests that human immortalized MSCs could be used for a large scale production of cells for mediated drug delivery. Moreover, the secretion of drug-associated MVs could represent a new way for producing new drug formulation by "biogenesis". In the context of the "advanced cell therapy procedure", the MVs/PTX production would use less resource and time and it could possibly contribute to simplification of GMP procedures.
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Via Pascal 36, 20133 Milan, Italy.|Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico, Maxillofacial and Dental Unit. Department of Biomedical, Surgical and Dental Sciences, University of Milan, Via Commenda 10, 20122, Milan, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Cocce
authors.affiliation: Medestea Research and Production Laboratories, Consorzio CARSO, Bari, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Balducci
authors.affiliation: CNR-Institute of Cell Biology and Neurobiology, via del Fosso di Fiorano, 64-00143, Rome, Italy.
authors.forename: Maria L
authors.initials: ML
authors.lastname: Falchetti
authors.affiliation: Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Pascucci
authors.affiliation: Laboratory of Clinical Pathology and Neurogenetic Medicine, Fondazione IRCCS Neurological Institute Carlo Besta, Via Celoria 11, 20133 Milan, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Ciusani
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Via Pascal 36, 20133 Milan, Italy.|I.R.C.C.S. Istituto Ortopedico Galeazzi; Via Riccardo Galeazzi 4, 20161 Milan, Italy.
authors.forename: Anna T
authors.initials: AT
authors.lastname: Brini
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Via Pascal 36, 20133 Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Sisto
authors.affiliation: Biology and Genetics Division, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11, 25123 Brescia, Italy.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Piovani
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, IRCCS Neurological Institute C. Besta, via Celoria 11, 20133 Milan, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Alessandri
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, IRCCS Neurological Institute C. Besta, via Celoria 11, 20133 Milan, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Parati
authors.affiliation: Institute of Biopathology and Regenerative Medicine (IBIMER), Biomedical Research Center, SAS University of Granada, Granada, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Cabeza
authors.affiliation: Department of Biomedical, Surgical and Dental Sciences, University of Milan, Via Pascal 36, 20133 Milan, Italy.
authors.forename: Augusto
authors.initials: A
authors.lastname: Pessina
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems; D004354:Drug Screening Assays, Antitumor; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D015394:Molecular Structure; D017239:Paclitaxel / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D013539:Simian virus 40 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D019098:Telomerase / Q000737:chemistry / Q000378:metabolism; D000093542:Gemcitabine
chemical_list: D000970:Antineoplastic Agents; D003841:Deoxycytidine; D049452:Green Fluorescent Proteins; D019098:Telomerase; D017239:Paclitaxel; D000093542:Gemcitabine
keywords: Mesenchymal stromal cells; drug delivery.; gemcitabine; immortalized MSCs; microvesicles; paclitaxel