Department of Hematology/Oncology, University Children's Hospital III, 60590 Frankfurt am Main, Germany.
Haematologica. 2010 Apr;95(4):651-9. doi: 10.3324/haematol.2009.015065. Epub 2010 Feb 23.
Background In vitro proliferative and differentiation potential of mesenchymal stromal cells generated from CD271(+) bone marrow mononuclear cells (CD271-mesenchymal stromal cells) has been demonstrated in several earlier and recent reports. In the present study we focused, in addition to proliferative and differentiation potential, on in vitro and in vivo immunosuppressive and lymphohematopoietic engraftment-promoting potential of these mesenchymal stromal cells compared to bone marrow-derived mesenchymal stromal cells generated by plastic adherence (plastic adherence-mesenchymal stromal cells).
We set up a series of experimental protocols in order to determine the phenotype of CD271-mesenchymal stromal cells, and their clonogenic, proliferative, differentiation and immunosuppressive potential. The potential of CD271-mesenchymal stromal cells to improve the engraftment of CD133(+) hematopoietic stem cells at co-transplantation was evaluated in immunodeficient NOD/SCID-IL2Rgamma(null) mice.
In vitro studies demonstrated that CD271-mesenchymal stromal cells differentiate along adipogenic, osteogenic and chondrogenic lineages (trilineage potential), produce significantly higher levels of cytokines than plastic adherence-mesenchymal stromal cells, and significantly inhibit the proliferation of allogeneic T-lymphocytes in mixed lymphocyte reaction assays. Elevated levels of prostaglandin E(2), but not nitric monoxide, mediated the majority of this immunosuppressive effect. In vivo studies showed that CD271-mesenchymal stromal cells promoted significantly greater lymphoid engraftment than did plastic adherence-mesenchymal stromal cells when co-transplanted with CD133(+) hematopoietic stem cells at a ratio of 8:1 in immunodeficient NOD/SCID-IL2Rgamma(null) mice. They induced a 10.4-fold increase in the number of T cells, a 2.5-fold increase in the number of NK cells, and a 3.6-fold increase in the number of B cells, indicating a major qualitative difference between these two mesenchymal stromal cell populations. Conclusions Our results indicate that CD271 antigen provides a versatile marker for prospective isolation and expansion of multipotent mesenchymal stromal cells with immunosuppressive and lymphohematopoietic engraftment-promoting properties. The co-transplantation of such cells together with hematopoietic stem cells in patients with hematologic malignancies may prove valuable in the prevention of impaired/delayed T-cell recovery and graft-versus-host disease.
在先前和近期的一些研究中，已经证明了从 CD271(+)骨髓单核细胞（CD271-间充质基质细胞）中生成的间充质基质细胞的体外增殖和分化潜能。在本研究中，我们除了关注增殖和分化潜能外，还重点研究了这些间充质基质细胞与通过塑料贴附生成的骨髓来源的间充质基质细胞（塑料贴附-间充质基质细胞）相比，其在体外和体内的免疫抑制和淋巴造血植入促进潜能。
我们建立了一系列实验方案，以确定 CD271-间充质基质细胞的表型及其克隆形成、增殖、分化和免疫抑制潜能。通过在免疫缺陷性 NOD/SCID-IL2Rgamma(null)小鼠中进行共移植，评估 CD271-间充质基质细胞改善 CD133(+)造血干细胞植入的潜能。
体外研究表明，CD271-间充质基质细胞可沿着脂肪、成骨和软骨谱系分化（三系潜能），产生的细胞因子水平明显高于塑料贴附-间充质基质细胞，并在混合淋巴细胞反应测定中显著抑制同种异体 T 淋巴细胞的增殖。大多数这种免疫抑制作用是由前列腺素 E2 介导的，而不是一氧化氮。体内研究表明，在免疫缺陷性 NOD/SCID-IL2Rgamma(null)小鼠中，当以 8:1 的比例与 CD133(+)造血干细胞共移植时，CD271-间充质基质细胞比塑料贴附-间充质基质细胞显著促进更大的淋巴系植入。它们诱导 T 细胞数量增加 10.4 倍，NK 细胞数量增加 2.5 倍，B 细胞数量增加 3.6 倍，表明这两种间充质基质细胞群体之间存在主要的定性差异。
我们的结果表明，CD271 抗原为分离和扩增具有免疫抑制和淋巴造血植入促进特性的多能间充质基质细胞提供了一种多功能标记物。在血液系统恶性肿瘤患者中，与造血干细胞共移植这些细胞可能有助于预防 T 细胞恢复受损/延迟和移植物抗宿主病。
