UNASSIGNED: Umbilical cord-derived MSCs (UC-MSCs) were gene-edited to avoid allogeneic immunity. The HLA class I expression was abrogated by the knock-out of the beta-2-microglobulin (B2M) gene; instead, the B2M-HLA-G fusion gene was knocked-in using the CRISPR/Cas9 system in combination with adeno-associated virus (AAV).
UNASSIGNED: Cell surface markers on gene-edited UC-MSCs were not different from those on primary UC-MSCs. The gene-edited UC-MSCs also retained the potential to differentiate into adipocytes, osteoblasts, and chondrocytes. B2M gene knock-out alone protected cells from allogeneic T cell immune responses but were vulnerable to NK cells. B2M gene knock-out in combination with B2M-HLA-G knock-in protected cells from both T cells and NK cells. The B2M-HLA-G knock-in MSCs retained a good immunosuppressive ability and the addition of these cells into the mixing lymphocyte reaction showed a significant inhibition of T cell proliferation.
UNASSIGNED: The results of this study demonstrated the possibility that the CRISPR/Cas9 system combined with AAV can be used to effectively disrupt/introduce any gene into UC-MSCs. Our findings suggest that the gene-edited cell line produced here using this method may have a higher ability to escape the cytotoxic activity of immune cells than primary cells, thereby being more advantageous for long-term graft survival.
未授权:对脐带来源的MSCs(UC-MSCs)进行基因编辑以避免同种异体免疫。通过敲除β-2-微球蛋白(B2M)基因来消除HLAI类表达;相反,使用CRISPR/Cas9系统联合腺相关病毒(AAV)敲入B2M-HLA-G融合基因.
未经鉴定:基因编辑的UC-MSCs上的细胞表面标记与原代UC-MSCs上的标记没有差异。基因编辑的UC-MSCs还保留了分化为脂肪细胞的潜力,成骨细胞,和软骨细胞.B2M基因敲除单独保护细胞免受同种异体T细胞免疫应答,但对NK细胞易感。B2M基因敲除与B2M-HLA-G敲入组合保护细胞免受T细胞和NK细胞两者的侵害。B2M-HLA-G敲入MSC保留了良好的免疫抑制能力,并且将这些细胞添加到混合淋巴细胞反应中显示出对T细胞增殖的显着抑制。
UNASSIGNED:这项研究的结果表明,与AAV结合的CRISPR/Cas9系统可用于有效破坏/将任何基因引入UC-MSC的可能性。我们的发现表明,使用这种方法产生的基因编辑细胞系可能比原代细胞具有更高的逃避免疫细胞细胞毒活性的能力,从而更有利于移植物的长期存活。