MSC Suppression Inspector (ヒト)
Description MSCは、繊維芽細胞様のプラスチック接着性の細胞で、骨髄細胞や脂肪組織など様々な組織から分離できます。近年になって、MSCの多分化能から免疫系調節機能へと、研究者の興味が移りつつあります。骨髄由来のMSCがT細胞の増殖を抑制することが報告されました1,2。このMSCの機能は、MSCの抑制能試験用に最適化されたT細胞刺激試薬を含む、MSC Suppression Inspectorで解析できます。このMSC Suppression Inspectorは、ビオチン標識されたCD2、CD3、CD28抗体が予めロードされたAnti-Biotin MACSiBead Particlesです。ポリクローナルな刺激（この場合はMSC Suppression Inspector）と共に、MSCと CD4+CD25– あるいは CD4+ responder T 細胞 (Tresp) と、異なる比率で共培養します。Trespは単独培養では増殖しますが、MSCと共培養するとTrespの増殖が悪くなります。細胞の増殖は 3H-チミジンで解析できますが、carboxyfluorescein succinimidyl ester (CFSE) ラベルによっても検出できます。 Applications ヒトMSCの、in vitro抑制試験による機能的な同定 Further information Stem Cell Research Brochure [PDF; 3,8 MB] Stem Cell Product List Sept 2011 [PDF; 709,8 KB]		Figure 1 MSCs were isolated from human bone marrow and culture expanded with NH Expansion Medium (# 130-091‑680). After two passages MSCs were co-cultured with CD4+CD25– responder T cells at different ratios. For T cell stimulation, the MSC Suppression Inspector was added to the culture. Proliferation of T cells was determined by 3H-thymidine incorporation. Tresp show high proliferation after stimulation with the MSC Suppression Inspector (A). When adding MSCs Tresp proliferation is suppressed dramatically (B). Unstimulated Tresp show no proliferation (D). MSCs alone show little proliferation with or without stimulation (C and E). Figure 2 MSCs were isolated from human bone marrow either by plastic adherence (PA-MSCs) or by CD271 isolation (CD271-MSCs) using CD271 MicroBead Kit (APC) (# 130-092-283). Both PA-MSCs and CD271-MSC were culture expanded with NH Expansion Medium (# 130-091‑680). After two passages MSCs were co-cultured with CFSE-labeled CD4+CD25– responder T cells. For T cell stimulation, the MSC Suppression Inspector was added to the cultures. The percentage of proliferating Tresp was measured as CFSE dye dilution analyzed by flow cytometry using the MACSQuant® Analyzer. Almost 90% of all Tresp proliferate after stimulation with the MSC Suppression Inspector (A). When adding PA-MSCs Tresp proliferation is suppressed to a level of about 28% (B). The addition of CD271+ MSCs suppresses the Tresp proliferation to a level of 20% (D). MSCs alone show no proliferation with or without stimulation (C, E, G, and H). Unstimulated Tresp show a proliferation rate of 30% (F).

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