title: Long-term stability, functional competence, and safety of microencapsulated specific pathogen-free neonatal porcine Sertoli cells: a potential product for cell transplant therapy.
abstract: BACKGROUND
  Porcine Sertoli cells (pSCs) have been employed for cell therapy in pre-clinical studies for several chronic/immune diseases as they deliver molecules associated with trophic and anti-inflammatory effects. To be employed for human xenografts, pSCs products need to comply with safety and stability. To fulfill such requirements, we employed a microencapsulation technology to increase pre-transplant storage stability of specific pathogen-free pSCs (SPF-pSCs) and evaluated the in vivo long-term viability and safety of grafts.
  
  
  METHODS
  Specific pathogen free neonatal pigs underwent testis excision under sterility. pSCs were isolated, characterized by immunofluorescence (IF) and cytofluorimetric analysis (CA) and examined in terms of viability and function [namely, production of anti-müllerian hormone (AMH), inhibin B, and transforming growth factor beta-1 (TFGβ-1)]. After microencapsulation in barium alginate microcapsules (Ba-MC), long-term SPF-pSCs (Ba-MCpSCs) viability and barium concentrations were evaluated at 1, 24 throughout 40 h to establish pre-transplant storage conditions.
  
  
  RESULTS
  The purity of isolated pSCs was about 95% with negligible contaminating cells. Cultured pSCs monolayers, both prior to and after microencapsulation, maintained high function and full viability up to 24 h of storage. At 40 h post-encapsulation, pSCs viability decreased to 80%. Barium concentration in Ba-MCpSCs lagged below the normal maximum daily allowance and was stable for 4 months in mice with no evident side effects.
  
  
  CONCLUSIONS
  Such results suggest that this protocol for the isolation and microencapsulation of pSCs is compatible with long-haul transportation and that Ba-MCpSCs could be potentially employable for xenotransplantation.
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.|Division of Medical Andrology and Endocrinology of Reproduction, Saint Mary Hospital, Terni, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Luca
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Mancuso
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Calvitti
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Iva
authors.initials: I
authors.lastname: Arato
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Falabella
authors.affiliation: Department of Pathology, Diagnostic and Clinical Veterinary Medicine, University of Perugia, Perugia, Italy.
authors.forename: Antonello
authors.initials: A
authors.lastname: Bufalari
authors.affiliation: Department of Pathology, Diagnostic and Clinical Veterinary Medicine, University of Perugia, Perugia, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: De Monte
authors.affiliation: Experimental Zooprophylactic Institute of Lombardia and Emilia Romagna, Brescia, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Tresoldi
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Nastruzzi
authors.affiliation: Department of Medicine, University of Perugia, Perugia, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Basta
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Fallarino
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Lilli
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Catia
authors.initials: C
authors.lastname: Bellucci
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Tiziano
authors.initials: T
authors.lastname: Baroni
authors.affiliation: Department of Medicine, University of Perugia, Perugia, Italy.
authors.forename: Maria Chiara
authors.initials: MC
authors.lastname: Aglietti
authors.affiliation: Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Giovagnoli
authors.affiliation: Department of Internal Medicine, University of South Florida, Tampa, FL, USA.
authors.forename: Don F
authors.initials: DF
authors.lastname: Cameron
authors.affiliation: Department of Experimental Medicine, University of Perugia, Perugia, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Bodo
authors.affiliation: Division of Medical Andrology and Endocrinology of Reproduction, Saint Mary Hospital, Terni, Italy.|Department of Medicine, University of Perugia, Perugia, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Calafiore
mesh_terms: D000464:Alginates; D000818:Animals; D000831:Animals, Newborn; D002469:Cell Separation; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D008297:Male; D051379:Mice; D012708:Sertoli Cells / Q000166:cytology / Q000502:physiology / Q000637:transplantation*; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D014183:Transplantation, Heterologous / Q000379:methods*
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid
keywords: Sertoli cells; microcapsules; specific pathogen free; xenotransplantation