Virally and physically transgenized equine adipose-derived stromal cells as a cargo for paracrine secreted factors.
BACKGROUND
Adipose-Derived Stromal Cells have been shown to have multiple lineage differentiation properties and to be suitable for tissues regeneration in many degenerative processes. Their use has been proposed for the therapy of joint diseases and tendon injuries in the horse. In the present report the genetic manipulation of Equine Adipose-Derived Stromal Cells has been investigated.


RESULTS
Equine Adipose-Derived Stromal Cells were successfully virally transduced as well as transiently and stably transfected with appropriate parameters, without detrimental effect on their differentiation properties. Moreover, green fluorescent protein alone, fused to neo gene, or co-expressed as bi-cistronic reporter constructs, driven by viral and house-keeping gene promoters, were tested. The better expressed cassette was employed to stably transfect Adipose-Derived Stromal Cells for cell therapy purposes. Stably transfected Equine Adipose-Derived Stromal Cells with a heterologous secreted viral antigen were able to immunize horses upon injection into the lateral wall of the neck.


CONCLUSIONS
This study provides the methods to successfully transgenize Adipose-Derived Stromal Cells both by lentiviral vector and by transfection using optimized constructs with suitable promoters and reporter genes. In conclusion these findings provide a working platform for the delivery of potentially therapeutic proteins to the site of cells injection via transgenized Equine Adipose-Derived Stromal Cells.
Donofrio
Gaetano
G
Dipartimento di Salute Animale, Sezione di Malattie Infettive, Facoltà di Medicina Veterinaria, via del Taglio 10, 43100 Parma, Italy. gaetano.donofrio@unipr.it
Capocefalo
Antonio
A
Franceschi
Valentina
V
Morini
Giorgio
G
Del Bue
Maurizio
M
Conti
Virna
V
Cavirani
Sandro
S
Grolli
Stefano
S
D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D003001:Cloning, Molecular; D005240:Feasibility Studies; D015316:Genetic Therapy; D048091:Guided Tissue Regeneration; D006736:Horses; D007109:Immunity / Q000235:genetics; D007114:Immunization; D007592:Joint Diseases / Q000473:pathology / Q000628:therapy*; D016180:Lentivirus Infections / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D017961:Lentiviruses, Equine; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods*
D000956:Antigens, Viral