Introduction: The application of stem cells in various medical fields is a well-known challenge for many researchers; hence the need to find more efficient cell sources for experimental manipulation as well as regenerative medicine and transplantation procedures. Tissues harvested from donors could represent an innovative and unlimited reservoir of different type of stem cells. Since tissues are routinely derived from cadaveric donors they can be used for the isolation of mesenchymal stromal/stem cell population (MSCs). In this study, we described the successful isolation, in vitro expansion and characterization of cadaver mesenchymal stromal/stem cells (hC-MSCs) derived by long-time frozen human post mortem arterial segments. Materials and Methods: hC-MSCs were obtained from explanted variously-sized arteries derived from post mortem donors after 4 days post mortem and following liquid nitrogen storage in tissue-banking facilities for more than 5 years. Immunophenotype, self-renewal and differentiation potential were determined using flow cytometry, in vitro assays, histological and immunological staining, molecular and ultrastructural approaches. Results: We successful isolated a cells population that survived to a prolonged ischemic insult and long-time cryopreservation. hC-MSCs appear with spindle-shape morphology, plastic adherent growth forming multiple confluence colonies and high proliferation rate. Phenotypically, they displayed numerous mesenchymal surface antigens (CD44, CD73, CD90, CD105, HLA), stemness (Stro-1, Oct-4, Notch-1), pericyte (CD146, PDGFR-β, NG2) and neuronal (Nestin) markers and lacked the expression of hematopoietic and vascular markers. In addition, these cells exhibited colony-forming ability using extreme cell seeding diluition; to form free floating embryoid-like bodies when grown in suspension; high immunosuppressive properties and multipotency for their ability to differentiate in multiple mesegenic lineages, i.e., adipogenic, osteo/chondrogenic, leiomyogenic and angiogenic. Discussion: Based on these results in addition to the easy accessibility, non-controversial, safety and abundant stem cell number, the procurement of plentiful hC-MSCs from cadaveric vascular tissues could be a scientific revolution in the field of cell therapy for many patients with severe diseases.

Cryopreserved post mortem arteries as an innovative cadaveric mesenchymal stem cell source

VALENTE, SABRINA;ALVIANO, FRANCESCO;CIAVARELLA, CARMEN;STELLA, ANDREA;PASQUINELLI, GIANANDREA
2013

Abstract

Introduction: The application of stem cells in various medical fields is a well-known challenge for many researchers; hence the need to find more efficient cell sources for experimental manipulation as well as regenerative medicine and transplantation procedures. Tissues harvested from donors could represent an innovative and unlimited reservoir of different type of stem cells. Since tissues are routinely derived from cadaveric donors they can be used for the isolation of mesenchymal stromal/stem cell population (MSCs). In this study, we described the successful isolation, in vitro expansion and characterization of cadaver mesenchymal stromal/stem cells (hC-MSCs) derived by long-time frozen human post mortem arterial segments. Materials and Methods: hC-MSCs were obtained from explanted variously-sized arteries derived from post mortem donors after 4 days post mortem and following liquid nitrogen storage in tissue-banking facilities for more than 5 years. Immunophenotype, self-renewal and differentiation potential were determined using flow cytometry, in vitro assays, histological and immunological staining, molecular and ultrastructural approaches. Results: We successful isolated a cells population that survived to a prolonged ischemic insult and long-time cryopreservation. hC-MSCs appear with spindle-shape morphology, plastic adherent growth forming multiple confluence colonies and high proliferation rate. Phenotypically, they displayed numerous mesenchymal surface antigens (CD44, CD73, CD90, CD105, HLA), stemness (Stro-1, Oct-4, Notch-1), pericyte (CD146, PDGFR-β, NG2) and neuronal (Nestin) markers and lacked the expression of hematopoietic and vascular markers. In addition, these cells exhibited colony-forming ability using extreme cell seeding diluition; to form free floating embryoid-like bodies when grown in suspension; high immunosuppressive properties and multipotency for their ability to differentiate in multiple mesegenic lineages, i.e., adipogenic, osteo/chondrogenic, leiomyogenic and angiogenic. Discussion: Based on these results in addition to the easy accessibility, non-controversial, safety and abundant stem cell number, the procurement of plentiful hC-MSCs from cadaveric vascular tissues could be a scientific revolution in the field of cell therapy for many patients with severe diseases.
Vascular Research in Italy
9
9
Valente S; Alviano F; Ciavarella C; Buzzi M; Stella A; Pasquinelli G
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/390148
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