Clinical-grade mesenchymal stromal cells (MSC) are usually expanded from bone marrow (BMMSC) or adipose tissue (ADSC) using processes mainly differing in the use of fetal calf serum (FCS) or human platelet lysate (PL). We aimed to compare immune modulatory properties of clinical-grade MSC using a combination of fully standardized in vitro assays. BMMSC expanded with FCS (BMMSC-FCS) or PL (BMMSC-PL), and ADSC-PL were analyzed in quantitative phenotypic and functional experiments including their capacity to inhibit the proliferation of T, B, and NK cells. The molecular mechanisms supporting T-cell inhibition were investigated. These parameters were also evaluated after pre-stimulation of MSC with inflammatory cytokines. BMMSC-FCS, BMMSC-PL, and ADSC-PL displayed significant differences in expression of immunosuppressive and adhesion molecules. Standardized functional assays revealed that resting MSC inhibited proliferation of T and NK cells, but not B cells. ADSC-PL were the most potent in inhibiting T-cell growth, a property ascribed to IFN-γ-dependent indoleamine 2,3-dioxygenase activity. MSC did not stimulate allogeneic T cell proliferation but were efficiently lysed by activated NK cells. The systematic use of quantitative and reproducible validation techniques highlights differences in immunological properties of MSC produced using various clinical-grade processes. ADSC-PL emerge as a promising candidate for future clinical trials
Clinical-grade mesenchymal stromal cells produced under various GMP processes differ in their immunomodulatory properties: standardization of immune quality controls
BIFARI, Francesco;RICCIARDI, Mario;KRAMPERA, Mauro
2013-01-01
Abstract
Clinical-grade mesenchymal stromal cells (MSC) are usually expanded from bone marrow (BMMSC) or adipose tissue (ADSC) using processes mainly differing in the use of fetal calf serum (FCS) or human platelet lysate (PL). We aimed to compare immune modulatory properties of clinical-grade MSC using a combination of fully standardized in vitro assays. BMMSC expanded with FCS (BMMSC-FCS) or PL (BMMSC-PL), and ADSC-PL were analyzed in quantitative phenotypic and functional experiments including their capacity to inhibit the proliferation of T, B, and NK cells. The molecular mechanisms supporting T-cell inhibition were investigated. These parameters were also evaluated after pre-stimulation of MSC with inflammatory cytokines. BMMSC-FCS, BMMSC-PL, and ADSC-PL displayed significant differences in expression of immunosuppressive and adhesion molecules. Standardized functional assays revealed that resting MSC inhibited proliferation of T and NK cells, but not B cells. ADSC-PL were the most potent in inhibiting T-cell growth, a property ascribed to IFN-γ-dependent indoleamine 2,3-dioxygenase activity. MSC did not stimulate allogeneic T cell proliferation but were efficiently lysed by activated NK cells. The systematic use of quantitative and reproducible validation techniques highlights differences in immunological properties of MSC produced using various clinical-grade processes. ADSC-PL emerge as a promising candidate for future clinical trials
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