In this study, we first developed an expansion protocol for human bone marrow derived-MSC (hBM-MSC) using two different supplements for culture media, i.e. fetal bovine serum (FBS) or human platelet lysate (hPL). Interestingly, hPL supplement was more effective than FBS in expanding MSC. Afterwards, we characterized MSC and confirmed their genome stability through karyotype analysis and real time-PCR. MSC were then assessed in vitro for their ability to acquire the anti-inflammatory phenotype necessary for avoiding immune rejection and modulating host immune effector cells. MSC priming with TNF-α and IFN-γ led to increased ability in preventing NK cell-mediated lysis. Moreover, using standardized proliferation assays, MSC displayed strong immune suppressive activity towards T, B and NK cells. We then obtained a reproducible xenogeneic mouse model of aGvHD that was used to assess in vivo the efficacy of hPL-expanded MSC-based immunotherapy with different schedules of MSC administration

Isolation, expansion and functional characterization of hPL-expanded hBM-MSC for the treatment of systemic and severe acute Graft-versus-Host Disease

Gatti Alessandro
2018-01-01

Abstract

In this study, we first developed an expansion protocol for human bone marrow derived-MSC (hBM-MSC) using two different supplements for culture media, i.e. fetal bovine serum (FBS) or human platelet lysate (hPL). Interestingly, hPL supplement was more effective than FBS in expanding MSC. Afterwards, we characterized MSC and confirmed their genome stability through karyotype analysis and real time-PCR. MSC were then assessed in vitro for their ability to acquire the anti-inflammatory phenotype necessary for avoiding immune rejection and modulating host immune effector cells. MSC priming with TNF-α and IFN-γ led to increased ability in preventing NK cell-mediated lysis. Moreover, using standardized proliferation assays, MSC displayed strong immune suppressive activity towards T, B and NK cells. We then obtained a reproducible xenogeneic mouse model of aGvHD that was used to assess in vivo the efficacy of hPL-expanded MSC-based immunotherapy with different schedules of MSC administration
2018
MSC, GvHD, IMMUNOTHERAPY
File in questo prodotto:
File Dimensione Formato  
Isolation, expansion and functional characterization of hPL-expanded hBM-MSC for the treatment of systemic and severe acute Graft-versus-Host Disease.pdf

Open Access dal 25/09/2020

Descrizione: Tesi Sperimentale di Dottorato
Tipologia: Tesi di dottorato
Licenza: Accesso ristretto
Dimensione 4.83 MB
Formato Adobe PDF
4.83 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/978782
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact